Istituto Superiore di Sanità

Magyar Tudományos Akadémia

Eötvös Loránd Tudományegyetem

Ambaciasta d’Italia in Ungheria

XII HUNGARIAN-ITALIAN SYMPOSIUM ON SPECTROCHEMISTRY:

ENVIRONMENTAL POLLUTION AND HUMAN HEALTH

Pécs, 23 – 27 October 2005

http://www.speciation-analysis.net/Public/Events/2005/02/14/1320.html

ABSTRACT BOOK

Edited by

Viktor Gábor Mihucz (a) and Gyula Záray (a, b)

(a) Joint Research Group of Environmental Chemistry of Hungarian Academy of Sciences

and Loránd Eötvös University

(b) Department of Inorganic and Analytical Chemistry, Loránd Eötvös University

ISBN 963 463 800 7

Budapest, 2005

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Honorary Committee

F. De Luigi (MAE)

E. Garaci (ISS)

I. Klinghammer (ELTE)

P. Medgyes (OM)

Scientific Committee

S. Caroli (ISS)

L. Campanella (UNIROMA)

G. Cicognani (CEI)

A. Conti (CNR)

F. Cubadda (ISS)

A. Fasulo (CNR)

R. Fuoco (UP)

Gy. Heltai (MTA-SM)

J. Hlavay (MTA-KKB)

G. Nagy (PTE)

J. Posta (MKE-ST)

B. Righetti (IEB)

I. Salma (ELTE)

M. Pánczél (EH)

Gy. Záray (ELTE, MTA-KKB)

Organising Committee

V. G. Mihucz (MTA-ELTE)

J. Pusztai (MTA)

E. Tatár (ELTE)

I. Virág (ELTE)

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Organised by

L. Eötvös University (ELTE), Budapest

Istituto Superiore di Sanità (ISS), Roma

Ambasciata d’Italia a Budapest

Hungarian Academy of Sciences (MTA), Budapest

Società Chimica Italiana, Divisione di Chimica Analitica

in cooperation with

Magyar Tudományos Akadémia Környezetkémiai Bizottsága (MTA-KKB), Budapest

Magyar Tudományos Akadémia Spektrokémiai Munkabizottsága (MTA-SM), Budapest

Oktatási Minisztérium (OM), Budapest

Pécsi Tudományegyetem (PE), Pécs

Università di Pisa (UP), Pisa

Consiglio Nazionale delle Ricerche (CNR), Roma

Ministero degli Affari Esteri (MAE), Roma

Central European Initiative (CEI)

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Sponsored by:

Miskolci Egyetem, Mechatronikai és Anyagtudományi Kooperációs Kutatási Központ

(MeAKKK)

Pécsi Tudományegytem, Dél-Dunántúli Kooperációs Kutatási Központ

(DDKKK)

HUNGAROPAAR Kft.

8200 Veszprém, Tüzér u. 71/I. Tel: (00-36-88) 590-390; Fax (00-36-88) 590-391

E-mail: info.hu@anton-paar.com http://www.anton-paar.hu/

UNICAM Magyarország Kft.

1144 Budapest, Kőszeg u. 29. Tel: (00-36-1)-221-5536; Fax: (00-36-1)-221-5531

E-mail: unicam@mail.datanet.hu http://www.kszgysz.hu/unicam.htm

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PREFACE The XII Hungarian – Italian Symposium on Spectrochemistry: Environmental Pollution and Human Health takes place at the Headquarters of the Hungarian Academy of Sciences in Pécs (Hungary), from 23 through 27 October 2005. The past events took place at Rome (1983), Budapest (1985), Ispra (1987), Veszprém (1989), Pisa (1991), Lillafüred (1993), Rome (1995), Debrecen (1997), Siena (1999), Eger (2001) and Venice (2003). The twelfth edition of this symposium – permanent forum, which already resulted in intense and fruitful scientific co-operations between Italian and Hungarian institutions and scientists – brings together spectroscopists from al over the world having an interest in obtaining reliable quantitative data for a better protection of human health and the environment. With about 90 presentations, an overview is presented in field of analytical chemistry from the point of view of food safety, environmental protection, biochemical studies, method innovation and instrumental development. The variety of the issues illustrated and the experimental approaches suggested testify the important information provided by this Symposium along with the progress made so far by the analytical sciences.

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PROGRAMME

OF THE XII HUNGARIAN – ITALIAN SYMPOSIUM ON SPECTROCHEMISTRY Monday, 24 October 2005 10:00 – 10:20 Opening of the Symposium and welcome addresses Oral session I Advances in new methodologies for environmental protection I Chaipersons: Klára Polyák and Sergio Caroli 10:30 – 11:00 A. Roda, D. Parisi, A. Zattoni, M. Magliulo, M. Guardigli, M.

Forina, P. Reschiglian: Analytical proteomics using soft impact mass spectrometry

11:00 – 11:30 A. Roda, M. Guardigli, M. Mirasoli, E. Michelini, M. Musiani: Molecular bio-chemiluminescence imaging

11:30 – 12:00 L. Campanella: Photosensors for quality and stability evaluation 12:00 – 12:30 Th. Schäfer, F. di Francesco, R. Fuoco: Microgravimetric sensors

based on ionic liquids 12:30 – 13:00 P. Fodor: Present and future of trace element analysis in food

control 13:00 – 14:00 Lunch break Oral Session II Environmental pollution and human health Chairpersons: Aldo Roda and Gyula Záray 14:00 – 14:30 P. Cescon, P. Campostrini, G. Capodaglio, A. di Domenico, R.

Fuoco, G. Venier, S. Caroli: Implementation of the decrees on the quality of water in the Venice lagoon

14:30 – 15:00 C. Turetta, G. Capodaglio, W. Cairns, S. Giuliani, M. Frignani, P. Cescon: Environmental Quality Assessment. Heavy Metal Distribution in the Tam Giang-Cau Hai Lagoon (Vietnam)

15:00 – 15:20 M. Óvári, H. Hoefler, C. Streli, Gy. Záray: Investigation of the carbon content of biofilms during the colonization period by total reflection X-ray fluorescence spectrometry

15:20 – 15:40 I. Rezić, I. Steffan: Impact of heavy metals from textiles on human health

15:40 – 16:00 Coffee break Oral Session III Advances in new methodologies for environmental protection II Chairpersons: Maria Perla Colombini and János Lakatos 16:00 – 16:30 E. Sabbioni: Analytical nanotoxicology: a new branch of analytical

chemistry in in vitro nanoparticles toxicology research for the protection of human health

16:30 – 17:00 J. Kristóf, E. Horváth, L. Vázquez Gómez, A. De Battisti: Spectrochemical investigation of electrode coatings designed for electrochemical waste water treatment

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17:00 – 17:20 J. Hassler, P. Barth, Gy. Záray, K. Florian: Analytical capabilities of ETV-ICP-OES method applied for analysis of ceramic, biological and environmental materials

Oral Session IV Trace elements, metabolites and human health Chairpersons: Enrico Sabbioni and Luigi Campanella 17:30 – 18:00 M. Patriarca, A. Menditto: Biomarkers – an effective tools to assess

human exposure to chemicals in everyday life 18:00 – 18:30 P. Bottoni: Emerging contaminants of water resources: pesticide

metabolites and pharmaceuticals 18:30 – 18:50 M. Zeiner, I. Steffan, F. Lintner, W. Schwägerl, P. Zenz: Influence

on elemental status by hip-endoprostheses 18:50 – 19:10 K. Zih-Perényi, A. Lásztity: Study of interaction between Pb and

pharmaceuticals by chelating sorbents and GFAAS Tuesday, 25 October 2005 Oral Session V Chemical speciation for environmental analysis Chairpersons: Luigi Campanella and György Heltai 9:00 – 9:30 L. Ebdon: The measurement of chromium speciation in

environmental pollution and its impact upon human health 9:30 – 9:50 L. Abrankó, Zs. Jókai, B. Kmellár, P. Fodor: Method development

for mercury speciation studies in fish samples intended for human consumption

9:50 – 10:10 V. Gergely, M. Montes-Bayón, R. Szalma, A. Sanz-Medel, P. Fodor: Speciation analysis of selenium in alfalfa sprouts by two-dimensional high-performance liquid chromatography with parallel ICP-MS and ESI-MS detection

10:10 – 10:30 E. Kápolna, M. Shah, J. A. Caruso, P. Fodor: Comprehensive selenium speciation studies in Se-enriched chives (Allium schoenoprasum) by HPLC- ICP-MS

10:30 – 10:50 R. Schaeffer, Cs. Soeroes, P. Fodor, W. Goessler, K. A. Francesconi, D. Kuehnelt: Arsenic speciation in Hungarian freshwater environment

10:50 – 11:10 M. Üveges, P. Rodríguez-González, P. Fodor, J. I. García Alonso: Isotope dilution analysis as a routine technique for analysing organotin compounds in seawater, sediment and biota samples

11:10 – 11:30 Coffee break Oral session VI Air pollution, environment and human health Chairpersons: Marina Patriarca and Les Ebdon 11:30 – 12:00 I. Salma, T. Labuda, W. Maenhaut, Gy. Záray: Contribution of

traffic-related sources to aerosol composition in downtown Budapest

12:00 – 12:20 S. Canepari, E. Cardarelli, M. Catrambone, C. Perrino, A. Pietrodangelo, M. L. Astolfi: A microanalytical procedure for the

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chemical characterisation of the inorganic content of atmospheric particulate matter

12:20 – 12:40 R. Ocskay, I. Salma, W. Maenhaut, Gy. Záray: Ionic composition of urban and rural background aerosols in Hungary

12:40 – 13:00 J. Osán, S. Török, B. Beckhoff, G. Ulm, C. Abete, R. Fuoco: Speciation of nitrogen compounds in nanoscopic fine aerosol samples using TXRF-NEXAFS

13:00 – 13:20 B. Berlinger, M. Náray: Comparison and evaluation of different sampling heads for welding fumes sampling

13:20 – 14:30 Lunch break Oral Session VII Biomonitoring and chemometrics for environmental analyses Chairpersons: József Posta and Roger Fuoco 15:00 – 15:30 G. Pepponi: Investigation of the correlation between pollen viability

and its elemental composition 15:30 – 16:00 A. Lengyel, K. Héberger, L. Paksy, O. Bánhidi, R. Rajkó: Evalution

of ground level ozone concentration using multivariate methods 16:00 – 16:30 K. Polyák: Dry and wet deposition of toxic metal into the Lake

Balaton 16:30 – 16:50 Z. Margitai, M. Braun, M. Leermakers: Statistical analysis of air

pollution in Romania using leaf samples 16:50 – 17:10 M. Braun, Z. Margitai, M. Leermakers: Tilia leaves as biomonitors

of air quality in Europe 17:10 – 17:30 István Papp, Mihály Braun: Riverine sediments as indicators of

pollution history 17:30 – 17:50 V. G. Mihucz, E. Tatár, I. Virág, L. Rácz, Gy. Záray: Determination

of rare earth elements in Hungarian wine samples and bentonites used for their purification by double focusing sector field inductively coupled plasma mass spectrometry

18:00 – 19:45 Poster Session (see detailed list at the end of the programme) Wednesday, 26 October 2005 Oral session VIII Speciation studies for environmental samples Chairpersons: Péter Fodor and Silvano Focardi 9:00 – 9:30 J. Posta, Á. Béni, R. Karosi, V. Andruch, J. Balogh: Separation of

chromium(VI) using complexation and its determination with GFAAS in environmental samples

9:30 – 9:50 E. Veschetti, D. Maresca, L. Lucentini, E. Ferretti, G. Citti, M. Ottaviani: Speciation analyses of vanadium in Etnean drinking water networks by SPE and ETAAS

9:50 – 10:15 J. Lakatos, V. Zs. Baranyai, A. Lengyel: Application of the speciation analysis based on sequential extraction in the waste recycling technologies

10:10 – 10:30 Á. Hargitai Tóth, Zs. Hartyáni, J. Balogh: Investigation of trace element distribution in unpolluted and polluted soil by sequential

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extraction procedure: particle size dependence 10:30 – 11:00 I. Varga: Iodine determination in dietary supplement products by

TXRF spectrometry 11:00 – 11:30 Coffee break Oral Session IX Anthropogenic nuclides in the environment Chairpersons: Gabriele Capodaglio and Jacob de Boer 11:30 – 12:00 M. Forte: The measurement of radioactivity in drinking water 12:00 – 12:20 P. Marcus, Z. Karpas, O. Paz-Tal, A. Lorber, H. Sela, Y. Hagag, L.

Salonen, H. Komulainen, A. Auvinen, H. Saha, P. Kurttio: The correlation between uranium in drinking water and its expression in bio-assays of urine, hair and nails

12:20 – 12:40 Zs. Varga, Zs. Stefánka: Rapid sequential methods for separation and pre-concentration of anthropogenic radionuclides followed by alpha-spectrometry and ICP-SFMS detection

12:45 – 13:30 Lunch break Oral Session X Molecular spectroscopy and chromatography for environmental

analyses Chairpersons: Géza Nagy and Maurizio Forte 13:30 – 14:00 Maria Perla Colombini: Archaelogical chemistry 14:00 – 14:30 J. de Boer, S. van Leeuwen, P. Korytar: Final results of the

European research project difference: New alternative methods for dioxin analysis

14:30 – 15:00 E. Horváth, K. Kristóf: Micro-Raman spectroscopic investigation of Lemna minor: implications for environmental monitoring application

15:00 – 15:30 S. Focardi, C. Guerranti, G. Perra, D. Baroni, I. Bisogno, T. Benincasa, A. Bianchini, A. Schiavone, N. Ademollo, F. Borghini: Levels of environmental pollutants in human blood and serum from Italy

15:30 – 16.00 E. Maloschik, E. Benfenati, A. Ernst, A. Székács: A monitoring program for pesticide contamination in surface waters in Hungary

16:00 – 16:20 J. Yao, A. Djah, J.-chan Wu, H.-yin Han, Gy. Záray: Analysis of some elements in four typical foods from Wuhan

20:00 Banquet Thursday, 27 October 2005 Oral Session XI Legislation and environmental analyses Chairpersons: Imre Varga and Gianfranco Pepponi 9:00 – 9:30 L. Bezur: RoHS Directive – How to do the related analysis 9:30 – 10:00 S. Caroli: The activities of the working group on good laboratory

practice of the organisation for economic co-operation and

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development 10:00 – 10:30 B. László, A. Zagyva, Gy. Heltai: Application of EU Water

Framework directives in monitoring of small water catchment areas in Hungary I. Selection and characterization of a reference site for small creeks in Mátra mountains

10:30 – 10:50 E. Szlepák, Gy. Heltai, E. Szilágyi, B. Csányi, E. Sz. Bardóczy:

Application of EU Water Framework directives in monitoring of small water catchment areas in Hungary II. Characterization of the moderately loaded, rural catchment area of Galga stream

10:50 – 11:10 G. Halász, I. Fekete, F. Szilágyi, B. Csányi, G. Heltai: Application of EU water framework directives in monitoring of small water catchment areas III. Characterisation of a heavily loaded urban area

11:10 – 11:30 K. Kristóf, A. Algaidi, Gy. Kampfl, Gy. Heltai: Influence of C/N ratio on NOx and CO2 production of cultivated soil in closed mesocosmos system

11:30 Closing ceremony

POSTER SESSION (TUESDAY, 26 OCTOBER 2005, 18:00 – 19:45) P01 A. Andreotti, M. P. Colombini, I. Degano, J. Lucejko: Colour fading in textiles:

An insight on degradation products P02 M. Orlandi, M. P. Colombini, F. Modugno, M. Sardelli, E-L. Tolppa, L. Zoia:

Archaeological wood characterisation by NMR and PY/GC/MS P03 M. P. Colombini, F. Modugno, E. Ribechini: Characterisation of vegetable oils in

archaeological findings P04 É. Bandi, L. Nagy, L. Sárga, G. Nagy: Effect of alkali metal content on wood ash

in fluid bed boiler P05 Á. Béni, I. Kiss, S. Berényi, J. Posta: Study of atomization and generating

processes with the FES/AAS combustion system ensuring 100% sample input P06 I. Kiss, L. Varga, J. Posta: Following of “in vitro” extraction of metals from tablets

by atomic absorption spectrometry P07 R. Kovács, Á. Béni, R. Karosi, Cs. Sógor, J. Posta: Determination of chromium

content in foodstuffs and nutrition supplements P08 G. Blo, A. Ceccarini, C. Conato, C. Contado, F. Dondi, F. Fagioli, R. Fuoco, A.

Pagnoni, A. Passerini: Development of SdFFF Coupled On-line to CID-ETAAS for the Characterization of Environmental Colloids

P09 F. Di Francesco, M. Onor, S. Tabucchi, M. G. Trivella, A. Ceccarini, T. Schäfer, R. Fuoco: Advances in clinical application of Breath analysis by gas chromatography/mass spectrometry

P10 R. Fuoco, S. Giannarelli, B. Muscatello, P. Bogani, M. Buiatti, A. Rosati: Screening of endogenous cytokinins and indole-3-acetic acid in Nicotiana glauca plants by chemical derivatization and gas chromatography-mass spectrometry analysis

P11 I. Marchetti, L. Menichetti, G. Tamborini, C. Kusmic, C. Cappelletti, L. Gaetano, G. Lazzerini, R. Fuoco, P. A. Salvadori, A. L’Abbate, M. Betti: Imaging elemental distribution in thin cryosections of soft tissues by SIMS

P12 M. Lugand Bourgeois, A. Cornely: Elimination of heavy metals intoxication. Sequential Micronutrition

P13 Cs. Csutorás, A. Kiss: An efficient method for the characterisation of the interaction of pesticides and different soil samples

P14 A. Detcheva, J. Hassler: Determination of Al, Ca, Cu, Fe, Mg, Mn, P and Zn in

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potato and grain by means of ETV-ICP-OES P15 M. I. Farkas, T. G. Weiszburg: Compositional investigation on settled and airborne

dusts with mineralogical methods P16 E. Ferretti, L. Lucentini, E. Veschetti, L. Bonadonna, A. Stammati, L. Turco, M.

Ottaviani: Many-sided strategy for screening and identification of unknown contaminants in water for human consumption

P17 E. Veschetti, B. Cittadini, M. Ottaviani, P. Leopardi, R. Crebelli: ICP-OES determination of vanadium in mice tissues after oral exposure

P18 Gy. Kampfl, K. Bálint-Kristóf, Gy. Heltai: Experimental microcosmos model for evaluation of N2O and CO2 production of cultivated soil

P19 A. Kiss, Cs. Csutorás: Investigation of the mechanism of photodegradation of different pesticides

P20 L. Mancini, P. Formichetti, E. Pierdominici, A. M. D’Angelo, M. Iaconelli, S. Fabiani, M. E. Beltrami, S. Marcheggiani, S. Larsen, G. Pace, L. Tancioni, P. Bottoni: Integrated approach for assessing ecosystem health

P21 C. Majdik, Á. Pénzes, I. Bartók, M. Tosa, P. Moldovan, F. Kilár, F. D. Irimie: Heavy metal monitoring in Phaseolus vulgaris in Transylvania

P22 Á. M. Móricz, Zs. Fatér, K. H. Otta, E. Tyihák: Overpressured layer chromatographic determination of aflatoxin B1, B2, G1 and G2 in red paprika

P23 Z. Murányi, Szabolcs B.Tóth: Redox reactions going on in wines P24 G. Muránszky, M. Óvári, Gy. Záray: Inductively coupled plasma-sector field mass

spectrometric determination of trace elements in the PM10 airborne dust fraction P25 M. Óvári, M. Zeiner, I. Steffan, Gy. Záray: Determination of platinum, palladium

and rhodium in human urine by electrothermal vaporization inductively coupled plasma sector field mass spectrometry (ETV-ICP-SFMS)

P26 L. Nagy, G. Nagy: Spectroscopic confirmation of electrocatalytic behavior of amperometric carbohydrate detection on copper electrode

P27 M. Pásztor T., G. Blazsó: Changes of brain water and ion concentration in experimental hypoxic brain oedema in rat

P28 I. Peltz-Császma, E. Andrási, A. Lásztity, P. Brätter: Cadmium and Zinc in Human Brain Samples

P29 E. Tatár, K. Mayer, T. L. Török, I. Virág, V. G. Mihucz, Gy. Záray: Determination of potassium isotope ratios by TIMS in biological fluids

P30 M. Zeiner, I. J. Cindric, I. Steffan: Trace elemental characterization of edible oils by ICP-AES and GFAAS

P31 K. Zih-Perényi, A. Lásztity: Preconcentration and GFAAS determination of vanadium in mineral waters and pharmaceuticals

P32 G. Cecchi, L. Mancini: Interaction between ecosystems and human health P33 F. Ferrara, D. Caronia, F. Fabietti, N. Ademollo, L. Mancini, E. Funari:

Alkylphenols and alkylphenol ethoxylates in wild fishes from the river Tevere (central Italy): a pilot study

P34 P. Formichetti, P. Andreani, L. Tancioni, C. Iacomini, L. Mancini: First data on the ecological characterization of the river Albegna (Tuscany, Italy)

P35 G. Pace, P. Formichetti, E. Pierdominici, L. Mancini: Macroinvertebrate and aquatic macropythes as indicators of freshwater quality

P36 L. Yi, Y. Chang-Ying, H. An-Xin, Q. Song-Sheng: Interaction between 2,2’-diselenadibenzoic acid and serum albumins investigated by fluorescence spectroscopy

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ABSTRACTS OF ORAL PRESENTATIONS

MONDAY, 24 OCTOBER 2005

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OL-1 ANALYTICAL PROTEOMICS USING SOFT IMPACT MASS SPECTROMETRY A. Roda1, D. Parisi1, A. Zattoni2, M. Magliulo1, M. Guardigli1, M. Forina3, P. Reschiglian2

1Dept. of Pharmaceutical Sciences, Via Belmeloro 6, I-40126 Bologna (Italy) 2Dept. of Chemistry “G. Ciamician”, Via Selmi 2, I-40126 Bologna (Italy), 3Dept. of Chemistry and Pharmaceutical and Food Technologies, Via Brigata Salerno, I-16147 Genoa (Italy) The term " proteomics" was coined to designate the large scale characterization of the entire protein complement of a cell line, tissue or organism. Mass spectrometry (MS) is today an indispensable tool for proteomic applications, which go from the "expression proteomics", to the study of multi-protein complexes or the "functional proteomics". The main driving forces for the development of biological MS have been novel ionization techniques able to transfer the biomolecules from the liquid phase to the gas phase, thus making them amenable to measurement in the mass spectrometer. The two dominant ionization methods are electrospray ionization (ESI) and matrix assisted laser desorption ionization (MALDI), which are defined "soft impact" ion sources. Common features of these methods include: generation of unfragmented protonated molecules, the ability to ionize biomolecules in the mass range of tens of kilodaltons, and successful analysis of a wide range of compounds in a variety of chemical classes. In the last few years MALDI and ESI sources were coupled with time of flight (TOF) analyzers because of the high resolution and mass accuracy of these analyzers, in the range of 10,000 and low ppm, respectively. Moreover new instrumentation for the analysis of the proteome has been developed such as MALDI and ESI hybrid quadrupole time of flight instruments (Q-TOF) which combine advantages of the mass finger printing and peptide sequencing methods for protein identification. In MALDI, ions arise from a solid phase, have low charge states, and can be formed from a sample containing many commonly used biological salts and buffers. In contrast, ESI ions are generated from solution, are formed with a range of charge states, and cannot be formed efficently in the presence of even small amounts of extraneous electrolytes. For this reason, sample preparation is a critical step to avoid risks of signal suppression due to competitive ionization in the presence of salts and contaminants. Proteins of interest can be purified using appropriate separation techniques, such as HPLC, or electrophoresis, which can be coupled on-line or off-line with the mass spectrometer. However, these separation techniques show limitations in the analysis of proteins in their native form, because of the interaction between proteins and the stationary phase, the presence of organic modifiers in the mobile phase, or the use of high voltages. Flow field-flow fractionation (FlFFF) is a flow-assisted technique particularly suited to the separation of intact proteins. No mechanical or shear stress on sample components is exerted since the fractionation channel does not contain a stationary phase. As mobile phase various carrier liquids can be used to avoid protein degradation. We have proposed the combination of an innovative, microchannel version of FlFFF, the hollow-fiber (HF) FlFFF, with TOF MS for the analysis of biological samples such as native proteins [1] and intact cells [2]. The intrinsic simplicity and low cost of our HF FlFFF version also allows for disposable usage. Disposable HF FlFFF can be particularly appealing for analytical bio-applications in which run-to-run repeatability is critical. To summarize, the analytical proteomic approach includes: 1) purification of protein from complex matrixs (i.e serum, cell lysate ecc), 2) MALDI-TOF or ESI-TOF MS analysis of the whole protein, 3) protein digestion in solution or in gel, 4) MALDI-TOF MS peptide mass fingerprinting for protein identification, 5) LC MS/MS

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analysis of peptides by using ESI-Q-TOF MS for de novo sequencing or for the identification of post-translation modifications. In this work we present some novel approaches to analytical proteomics based on soft impact mass spectrometry for the analysis of functional protein drugs and pathogenic bacteria. First we discuss an application in the evaluation of the purity profile and the detection of the enzymatic activity of a recombinant protein drug, the enzyme urate oxidase. The approach is based on a new combination of techniques: HF FlFFF, MALDI/TOFMS, and chemiluminescence (CL) enzyme assay. The combined approach follows the preliminary detection and identification of sample impurities by means of conventional methods: RP HPLC with ESI/Q-TOF MS, and MALDI/TOFMS with SDS PAGE and 2D SDS PAGE. Results show that uricase preparations obtained from recombinant microorganisms contain proteins from the host system, and that different uricase oligomers may be present with different enzymatic activity. The second study aims to the identification of pathogenic bacteria through fingerprinting protein expression profiles by MALDI-TOF MS of whole cells. Three different bacterial strains were analyzed: Escherichia coli MC 1061; Escherichia coli O157: H7; Yersinia enterocolitica at different growth time (4, 6, 9, 15, 18, 24 hours). Moreover different strains of Escherichia coli O157, were analyzed. In order to evaluate the difference in the protein expression profile of these bacteria, mass spectra were analyzed using different chemometric techniques such as the principal component analysis (PCA) or the linear discriminant analysis (LDA). This approach proved able to discriminate bacterial cultures of different species and at different growth time. [1] Reschiglian, P.; Zattoni, A.; Roda, B.; Cinque, L.; Parisi, D.; Roda, A.; Dal Piaz, F.; Moon, M.H.; Min, B.-R. Anal. Chem. 2005, 76, 47. [2] Reschiglian, P.; Zattoni, A.; Cinque, L.; Roda, B.; Dal Piaz, F.; Roda, A.; Moon, M.H.; Min, B.-R. Anal. Chem. 2004, 76, 2103.

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OL-2 MOLECULAR BIO-CHEMILUMINESCENCE IMAGING A. Roda1, M. Guardigli1, M. Mirasoli1, E. Michelini1, M. Musiani2

1Dept. of Pharmaceutical Sciences, Via Belmeloro 6, I-40126 Bologna (Italy) 2Dept. of Experimental and Clinical Medicine, Via Massarenti 9, I-40138 Bologna (Italy) In recent years, luminescence imaging techniques employing highly sensitive detection principles and probes have been successfully exploited for the development of analytical methods for the ex vivo and in vivo evaluation of the spatial and temporal distribution of target molecules involved in biological and pathological processes. Here recent progresses in luminescence imaging techniques are described, including new labels for fluorescence (FL), time-resolved fluorescence (TRF) and bioluminescence (BL), multiplexed imaging microscopy techniques relying on the combination of different detection principles and whole-body BL imaging in live animals. Fluorescence imaging techniques employing biospecific probes labeled with fluorophores are well-established bioanalytical tools in life sciences, and a wide range of efficient fluorophores with different spectral properties are commercially available. However, most of them present small shifts between excitation and emission wavelengths (Stokes’ shift), resulting in interference between the excitation and emitted lights. In addition, they have different optimal excitation wavelengths, which may not match the emission spectrum of the excitation source. To overcome these drawbacks, we have developed a BODIPY derivative (BODIPYs are well-known FL labels) with additional chromophoric groups [1], which is strongly fluorescent upon excitation at 370 nm, a wavelength compatible with mercury arc lamps or nitrogen lasers. This molecule, which also contains a suitable reactive group for coupling to biomolecules, proved suitable for fluorescent labelling of proteins. This approach could be extended to other BODIPYs to obtain a family of fluorophores for protein labelling with different emission spectra that can be excited at the same wavelength, thus suitable for the development of multiplexed FL imaging techniques. Interference due to scattered excitation light, as well as to sample background emission, can be also avoided by means of TRF techniques. In such techniques a FL label – typically a lanthanide chelate - with long luminescence lifetimes (micro- to milliseconds) is excited by a pulsed light source and the resulting emission is recorded after a suitable delay (typically a few hundreds of microseconds). Such delay allows for vanishing of the short-living sample background emission, while the label emission remains still detectable. A new label for TRF imaging was developed by synthesizing a ligand for lanthanide ions containing 2,2’-bipyridine-6-carboxylate units [2]. Such ligand gave stable and strongly luminescent europium and terbium complexes characterized by long luminescent lifetimes, which were applied to TRF imaging microscopy using a model label-BSA conjugate. In the last years BL techniques have also gained a relevant importance in bioanalysis and in general in life sciences due to the availability of various self-luminescent organisms and the possibility to obtain BL in a given cell in response to a specific stimulus or analyte through genetic engineering (reporter gene technology). Firefly, Renilla, and bacterial luciferases are among the most widely used BL proteins for these applications. However, the performance of BL bioanalytical systems could be improved by employing new BL proteins with higher thermostability, wider optimal pH ranges and improved emission properties. As an example, red- and green-emitting luciferases for applications such as in vivo imaging and multiplexed

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assays have been recently developed by site-directed and random mutagenesis of Photinus pyralis firefly luciferase [3]. The peculiar characteristics of BL/CL detection (high detectability and specificity, possibility to perform an easy quantitative evaluation of the BL/CL signal) have been also exploited in immunohistochemistry (IHC) and in situ hybridization (ISH) techniques for the sensitive localization and quantitation of the target analytes through the employment of enzyme-labeled biospecific probes. We have developed a CL IHC technique for the quantitative analysis of the Multidrug Resistant Protein 2 (MRP2) in liver biopsy sections [4], which allowed reliable and objective evaluation of the protein content in the tissue. When applied to clinical samples from patients with primary cirrhosis under therapy with ursodeoxycholic acid, it showed an increase of the MRP2 levels, in line with that reported in the literature and obtained with Western blotting or real-time PCR techniques. Multiplexed IHC and ISH imaging, usually performed by means of FL imaging techniques, can be also developed by combining different luminescence detection techniques. We have employed a combined ISH-IHC method for the co-localization of mucosal human papillomaviruses (HPVs) and the melanocitic marker HMB-45 in melanoma biopsy specimens [manuscript in preparation]. The method combined the chemiluminescent (CL) ISH detection of the HPV genome and the FL IHC localization of HMB-45. The two signals are separately triggered and acquired, then the resulting FL and CL images are digitally combined to compare the localization of the target species. The method was able to show the co-localization of both markers in the vast majority of cells, thus supporting the hypothesis of the involvement of HPV in skin cancer development and progression. The recent advancements in the BL reporter gene technology has made possible the development of BL whole-body cellular and molecular imaging techniques, which offered a sensitive, quantitative, non-invasive, and real-time method to monitor many biochemical and pathological processes in living animals, such as cancer growth and metastasis, infectious states and gene expression patterns. These techniques are nowadays used in animal models to perform in vivo efficacy, pharmacokinetics, toxicology and target validation studies, even in a high-throughput screening (HTS) format. We used in vivo low-light imaging of a BL recombinant Escherichia coli strain in a rat model to monitor the motion of the bacteria through the gastrointestinal tract [5]. Bacteria were administered by gavage and the light signal spatial and temporal distribution was evaluated by using a CCD-based low-light imaging detector at different time intervals, obtaining information without sacrificing animals at each data collection. In conclusion, luminescence imaging, especially CL and BL imaging techniques, represents powerful bioanalytical tools for life sciences, especially for the highly sensitive detection and localization of target molecules and the real-time in vivo monitoring of biochemical processes. The development of new BL/CL labels and proteins and the advancements in imaging technology are expected to lead in the next future to wider application of these techniques. [1] Ulrich G., Goze C., Guardigli M., Roda A., Ziessel R., Angew. Chem. Int. Ed. 44 (2005) 3694. [2] Weibel N., Charbonnière L.J., Guardigli M., Roda A., Ziessel R., J. Am. Chem. Soc. 126 (2004) 4888. [3] Branchini B.R., Southworth T.L., Khattak N.F., Michelini E., Roda A., Anal. Biochem. 345 (2005) 140. [4] Guardigli M., Marangi M., Casanova S., Grigioni W.F., Roda E., Roda A., J. Histochem. Cytochem. (in press, doi:10.1369/jhc.5A6621.2005). [5] Roda A., Guardigli M., Pasini P., Mirasoli M., Musiani M., Anal. Chim. Acta 541 (2005) 25.

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OL-3 PHOTOSENSORS FOR QUALITY AND STABILITY EVALUATION Luigi Campanella Department of Chemistry, University La Sapienza, Piazzale Aldo Moro 5, 00185 Rome (Italy) This is a preliminary approach to the use of a new generation of solid state sensors based on the capacity of the sensor element to catalyze the photodegradation of various kinds of organic compounds and to recognise their structure on the basis of the type of process catalyzed. The electron holes present in the TiO2 structure are able to trigger and oxidative process involving substances present in the environment, in particular those ones that can be adsorbed on it. This characteristics make titanium dioxide a suitable material for use as a sensor for antioxidant, i.e. reducing, substances. By reacting with it these substances produce potential variations corresponding to values specific of the above mentioned photo-oxidative processes. In this first attempt, TiO2 in the prevailing anatase form was used as sensor, and phenols and wine as known compounds and matrixes. The overall profile of the response, curve E = f(t), is diagnostic for the purpose of compound recognition, the characterization of its state purity or alteration or adulteration. The behaviour of nanoparticulated TiO2 was tested in the presence of antioxidants (hydroquinone, ascorbic acid, n-propylgallate) as well as of highly antioxidant complex matrixes (DOC wine, commercial wine, homemade wine). Interesting results have been obtained using real matrixes: for example, in the case of red wine, it was found that higher is the quality of the wine faster the onset of the signal after contact is made between the medium and the sensor. Lastly it was observed that the sensor responds satisfactorily also in the presence of adulteration of the real matrix. This adulteration was simulated by adding to the wine known volumes of methanol and was revealed by the anomalous potential jumps and shorter onset times observed compared with those observed for the genuine wine sample. In conclusion several experimental details were thoroughly studied and optimized: they include nanoparticulated titanium dioxide in suspension, the presence in the water used as solvent of carbonate and organic substances (which affect sensor response), the buffering properties of the phosphate over the entire linear range of detector response, the verification using alternative techniques of the results obtained, the rigorous analysis of the data (kinetic curves, uncertainty and equation describing the phenomenon); lastly, the size (owing to the growing demand for ‘portable’ instruments), ease of use and the materials characterizing the sensor.

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OL-4

MICROGRAVIMETRIC SENSORS BASED ON IONIC LIQUIDS

Thomas Schäfer1,&, Fabio Di Francesco2, Roger Fuoco1 1Dipartimento di Chimica e Chimica Industriale, Università di Pisa, Via Risorgimento, 35, 56126 Pisa 2Istituto di Fisiologia Clinica, CNR, Via Moruzzi, 1, 56100 Pisa &mailing address: Istituto di Fisiologia Clinica, CNR, Via Moruzzi, 1, 56100 Pisa, E-mail: tom@ifc.cnr.it, Fax: +39 050 315 21 66

Room temperature ionic liquids (RTILs) are salts, which are liquid at or near ambient

temperature. Because these liquids consist entirely of ions, practically they do not possess any measurable vapour pressure and hence do not significantly evaporate, even at an elevated temperature (limited by the temperature of decomposition). Although electrolytes in nature, they can possess a dissolution power similar to that of common organic solvents. Owing to these unique properties, RTILs have been investigated extensively recently as alternative non-polluting solvents in chemical processing. RTILs can to some extent be tailor-made for a specific application, such as a maximum affinity for target solutes, which is why they are commonly designated “designer solvents”. Their possible utilisation for sensor applications has only been recognized recently. RTILs composed of different cations and anions were deposited on the quartz resonator and its response to model solutes at varying concentrations in the vapour phase monitored. The resulting frequency shifts were positive, contrary to what is commonly observed with polymeric depositions and presumably owing to the viscosity of the ionic liquid decreasing upon dissolution of solutes.

The response of two ionic liquids, BMIM+PF6- (butylmethylimidazolium hexafluoro

phosphate) and BMIM+NTf2- (butylmethylimidazolium (bis((trifluoromethyl)sulfonyl)imide)

to water, ethanol and ethyl acetate, was studied. Both the steady-state and the transient response were investigated, as the latter yields indispensable information for discriminating between solutes. It was found that while BMIM+PF6

- yielded very similar frequency shifts for these three model solutes, BMIM+NTf2

- gave a far more pronounced response to ethanol, demonstrating how the selectivity of an ionic liquid can to some extent be tuned for target compounds. As for the transients, it could be seen that while in the same ionic liquid the diffusion velocity of solutes of different molar volumes is significantly different, it was found to be almost equal for the same compounds in different ionic liquids. The unique properties of the ionic liquids, namely the lack of a measurable vapour pressure, were furthermore explored in particular when using microgravimetric sensors under reduced pressure. The results obtained demonstrate that ionic liquids are versatile and represent a fascinating alternative for selectively interacting with target solutes in sensor devices. Their potential, but also the current limits of their application, will be highlighted.

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OL-5 PRESENT AND FUTURE OF TRACE ELEMENT ANALYSIS IN FOOD CONTROL Péter Fodor Corvinus University of Budapest, Department of Applied Chemistry, H-1118 Budapest, Villányi ut 29-33, Hungary The present situation of trace element analysis in food control has not changed in the past 100 years. The main philosophy of the routine analysis is to make efforts to report the total elemental concentrations of different food samples. The last 25 years, when the elemental speciation verified the different toxicity and different biological role of elemental species, still not a routine technique in the food analysis. On the other side there are several other speciation needs, not only in case of well investigated arsenic, selenium, mercury or antimony, but in case of zinc, iron, calcium or manganese where for the food quality, food safety or food technology decision, speciation analysis required. Examples from every day food analysis will be describe the new line of trace element analysis in food sector.

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OL-6 IMPLEMENTATION OF THE DECREES ON THE QUALITY OF WATER IN THE VENICE LAGOON P. Cescon1, P. Campostrini2, G. Capodaglio1, A. di Domenico3, R. Fuoco4, G. Venier2, S. Caroli3 1Ca’ Foscari University, Department of Environmental Sciences, Dorsoduro 2137, 30123 Venice, Italy. E-mail: cescon@unive.it; capoda@unive.it. 2Consortium for the Management of the Coordination Centre of Research Activities on the Venice Lagoon System, Palazzo Franchetti, S. Marco 2847, 30124 Venice, Italy. E-mail: campostrini@corila.it; venier@corila.it. 3Istituto Superiore di Sanità, Rome, Viale Regina Elena 299, 00161 Rome, Italy. E-mail: caroli@iss.it; addeke@iss.it. 4University of Pisa, Department of Chemistry and Industrial Chemistry, Via Risorgimento 35, 56126 Pisa, Italy. E-mail: fuoco@dcci.unipi.it. In recent years a number of legal provisions have been developed to monitor and ameliorate the quality of water in the lagoon of Venice. Known as the Ronchi-Costa Decrees, these acts were issued over the period 1998-1999 and dictate the maximum tolerable concentrations that potentially toxic and ecotoxic substances should not exceed both in the lagoon and in its adjoining basin. The decrees envisage a stepwise approach to reach the planned quality targets in the years 2008 and 2015 so as to progressively abate the chemical risk in the lagoon. The regulated contaminants include chemical elements [Ag, Al, As, B, Ba, Be, Cd, Co, Cr (total), Cr (trivalent), Cr (hexavalent), Cu, Fe, Hg, Mn, Mo, Ni, Pb, Sb, Se, V and Zn], inorganic substances (total nitrogen and total phosphorus as well as their various forms, bromate, residual chlorine, chloride, chlorite, cyanide, organochlorine compounds, fluoride, sulfide, sulfite, sulfate), anionic and non-ionic surfactants, phenols, halogenated organic solvents, aromatic organic compounds, polycyclic aromatic hydrocarbons, organochlorine and organophosphate pesticides, herbicides, dioxins and tributyltin. Parameters such as temperature, colour, smell, coarse and particulate matter, COD, BOD and Escherichia coli are also taken into account. The complexity of the tasks entrusted to monitoring authorities as well as to laboratories involved in the analytical activities calls for significant investments in terms of instrumentation and expertise. In this context, a Handbook has been recently set up in order to assist the experimentalist in meeting the stringent requirements prescribed by the said Decrees. As regards future regulatory activities in this field and the ensuing development of fit-for-purpose analytical methods, chemical speciation on the one hand and residues of pharmaceuticals and metabolites of pesticides on the other hand are emerging priorities to which the greatest attention should be devoted.

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OL-7 ENVIRONMENTAL QUALITY ASSESSMENT. HEAVY METAL DISTRIBUTION IN THE TAM GIANG-CAU HAI LAGOON (VIETNAM) Clara Turetta1, Gabriele Capodaglio1,2, Warren Cairns1, Silvia Giuliani3, Mauro Frignani3, Paolo Cescon1,2

1CNR, Institute for the Dynamics of Environmental Processes, Dorsoduro 2137, 30123 Venice (Italy) 2 University of Venice, Department of Environmental Science, Dorsoduro 2137, 30123 Venice (Italy) 3CNR, Institute of Marine Sciences-Section of Marine Geology, Via Gobetti 101, 40129 Bologna (Italy)

The contamination of aquatic systems, understanding the mechanisms of distribution

and accumulation, identifying the sources and trends may be achieved through the study of sediment and water: sediments can be considered as a record of environmental conditions at the time of their deposition; water provides information about current conditions and the mobility of pollutants. The paper presents research carried out in the Tam Giang-Cau Hai (TG-CH) Lagoon, located in the province of Thua Thien-Hué (central Vietnam). Although this lagoon is the largest in south east Asia, very few data are available about the presence of contaminants. In particular heavy metals represent an important class of contaminants that may derive from a number of natural sources and/or human activities.

The purpose of this work was to obtain information about the distribution of heavy metals in the lagoon water (Ag, As, Cd, Cr, Ni, Pb, U, V and Zn) and their distribution and accumulation in sediments. Measurements in lagoon water were performed using an ICP-SFMS (Element2, Finnigan-MAT, Germany) coupled with a desolvation introduction system. Measurements in sediment were performed using an ICP-QMS (Agilent 7500) fitted with a standard double-pass spray chamber and a v-groove nebulizer. Concentrations in the lagoon water (minimum–maximum in ng mL-1) were: Ag 0.00003-0.00614, As 0.21-1.61, Cd 0.01-0.12, Cr 0.04-0.27, Ni 0.04-4.43, Pb 0.04-0.37, U 0.17-2.04, V 0.35-1.56, Zn 0.01-1.69. Concentration ranges in the sediment (minimum–maximum in µg g-1) were: Ag 0.06-0.15, As 3.49-21.4, Cd 0.02-0.46, Cr 5.18-59.5, Ni 4.25-26.3, Pb 3.04-29.5, U 0.44-4.66, V 8.94-69.0, Zn 11.0-82.5.

The northern lagoon is characterized by the input of large amounts of fresh water from the O Lau River and presents a relatively high concentration of Ag and Cd in the sediment and low in the water column, while Cr and Ni featured an inverse partition between water sediment and water. The central part of the lagoon is characterized by the input of the Huong River, which goes through Huè City, and by the connection with open sea. This area featured the maximum concentration of As, Cr and Ni in sediment and the highest concentration of Ag, As, Cd, Cr, Ni, Pb in the water, suggesting a contribution from urban area of Huè. The southern part of the lagoon presents an irregular connection with sea and input from the Truoi River. It shows different partitions of trace elements between sediment and water with a trend related to the distance from the central lagoon. We can hypothesise that estuarine-like processes control the heavy metal distribution and partition in the northern lagoon as emphasized by the relation between water/sediment composition and salinity. The distribution of heavy metal in the southern lagoon seems to be controlled by the sedimentation and, as emphasized by the contrasting distribution between water and sediment, also by estuarine processes.

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The presence of relatively high metal concentrations in sediments seems to be related to the inputs of the rivers that flow into the lagoon. However, the distribution of the maximum values and the different water/sediment partition suggest diverse influences from the catchment areas. We can conclude that the TG-CH Lagoon is only slightly contaminated by heavy metals and only the As reaches values that could occasionally cause adverse effects on the most sensitive organisms.

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OL-8 INVESTIGATION OF THE CARBON CONTENT OF BIOFILMS DURING THE COLONIZATION PERIOD BY TOTAL REFLECTION X-RAY FLUORESCENCE SPECTROMETRY

Mihály Óvári1, H. Hoefler2, Christina Streli2, Gyula Záray3,4,5

1Eötvös University, Institute of Chemistry, Dept. of Chemical Technology and Environmental Chemistry, H-1518 Budapest, P.O.Box 32. 2Atominstitut of the Austrian Universities, TU-Wien, A-1020, Vienna, Austria. 3Eötvös University, Institute of Chemistry, Dept. of Inorganic and Analytical Chemistry, H-1518 Budapest, P.O.Box 32. 4Joint Research Group of Environmental Chemistry of the Hungarian Academy of Sciences & Eötvös University, H-1518 Budapest, P.O.Box 32. 5Hungarian Satellite Centre of Trace Elements Institute to UNESCO, H-1518 Budapest, P.O.Box 32. Biofilms (periphyton communities) are complex associations consisting bacteria, algae, funghi and their secretions, building up a mucous cover on all immersed surfaces in the aquatic environment. Since the biofilms are important nutrition for many organisms in the aquatic ecosystem, the trace elements bound by them can be directly introduced into the food chain. For the complex understanding of the biofilm formation and to judge better the effects of pollution on biofilms, it is necessary to determine not only the polluting trace elements but also the major biogenic elements (C,N,P,S) in biofilms. For element determination in minute biological samples, total reflection X-ray fluorescence spectrometry (TXRF) is very suitable. However, commercial spectrometers fitted with Mo-anode X-ray tube and air beam pathways can not be used for determination of low Z elements (Z<13). Using a soft X-ray source, airless beam pathways ans special detectors, this range can be extended down to Z=6, making it possible to determine elements as e.g. carbon. Recently, a new method has been developed for TXRF determination of low Z elements in biofilms directly cultivated on the quartz carrier plates. This method uses a Cr-anode X-ray tube and monochromatic X-ray primary beam with 5 keV energy. The internal standardization was made in two steps: 100 ng Ag was pipetted onto the plate, and the natural Ti content was determined over Ag. Then, natural Ti was used for the calculation of the carbon conent of the biofilm sample. The samples were obtained by cultivation in the Velence Lake (50 km SW from Budapest, Hungary). The cultivation time was 3, 6, 7, and 11 days. At each sampling two or three plates were taken to obtain data about the reproducibility. The plates were carefully washed by some drops od distilled water and cooled to +4 oC and dried at this temperature. On the basis of the results it can be established that this type of biofilm cultivation can be used for monitoring purposes. It is suggested to carry out the direct exposition of the plates not longer than about 6 - 7 days. The increase of the carbon content during cultivation is well determinable, however, after one week, the inhomogeneity increased considerably. The reason for this could be the non-equal distibution of large deposits visible on the surface. A longer cultivation is not recommendable, because of the increasing thickness of the biofilm layer, resulting in higher background in the spectrum.

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OL-9 IMPACT OF HEAVY METALS FROM TEXTILES ON HUMAN HEALTH Iva Rezić1, Ilse Steffan2

1Laboratory of Analytical Chemistry, Department of Textile Chemistry and Material Testing, Faculty of Textile Technology, University of Zagreb, Croatia 2Department of Analytical Chemistry and Food Chemistry, University of Vienna, Austria

Due to daily contact with textiles of clothes, bed linen, and similar products it is very important that those materials do not contain health–hazardous chemicals. Such chemicals belong to different groups: dyes, carriers, softeners, formaldehydes, heavy metals, biocides, and fire-retardants. Heavy metals are used in a variety of textile processing techniques. This includes oxidisation of vat and sulphur dyes, after treatment of direct dyes, catalytic reactions in curing resins, finishing, and dye stripping. All of the six common dye classes (acid, basic, direct, disperse, fibre reactive and vat) contain metals including chromium, arsenic, cadmium, mercury, copper, lead, and zinc. Raw textile materials may also contain larger quantities of heavy metals due to adsorption from the environment (e.g. from the air, water and soil).

A majority of problems arise in the field of heavy metal complex dyes being extracted from the fabrics by perspiration or saliva solutions. Therefore our primary target of interest was the determination of extracted heavy metals from different textile materials. Experiments were performed according to the Öko Tex standard for textile products used in direct contact with the human body. Leaching of heavy metals from textiles was carried out in a synthetic saliva solution. The artificial acidic sweat solution applied was prepared according to the ISO 105 – E04. After extraction from Croatian textile products made of cotton, flax, wool, silk, viscose, and polyester materials, heavy metals were determined by inductively coupled plasma – optical emission spectrometry (ICP-OES).

Although the results confirmed that in most cases the concentration of heavy metals did not exceed the permissible values, in some cases the concentrations of particular heavy metals were above the limits given by Öko Tex. This indicates that certain textile products may represent a health hazard to consumers and can cause skin irritations and allergies. They can also be a potential source of pollution in cases when the wastewaters from households, laundries or textile industries are being discharged into the environment.

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OL-10 ANALYTICAL NANOTOXICOLOGY: A NEW BRANCH OF ANALYTICAL CHEMISTRY IN IN VITRO NANOPARTICLES TOXICOLOGY RESEARCH FOR THE PROTECTION OF HUMAN HEALTH E. Sabbioni European Commission, ECVAM Unit, IHPC, JRC-Ispra, 21020 Ispra, Varese (Italy)

Nanotechnology, the science of the “infinitely small”, is considered as the key technology of the 21st century that have a “revolutionary” potential in terms of contributions in solving global environmental challenges. Currently, enormous progress is being made in producing a great number of nanoparticles (NP) and nanomaterials. However, in spite of hundreds of nanotechnology applications are already in commercial production a huge health and safety questions remain unsolved. There are good reasons to fear that manufactured NP could have undesirable health effects on general populations and occupationally exposed workers. Nanosized particles could be harmful to health due to their nanoscale (one billionth of meter) which even makes them possible to enter body cells crossing biological barriers such as blood-brain-barrier, the toxicity increasing as the size of the particle decrease. Unfortunately, in spite of the production and use of NP is becoming part of every-day life there is very little knowledge on the health impact of NP. In order to avoid the development of nanomaterials and then see whether they are dangerous from health point of view and take the measures to avoid negative health effects that have already happened scientific investigations, including toxicological experiments, must be urgently carried out to ensure that such particles are safe. In this context, recent studies in our laboratory suggest that the issues of NP toxicology research is more complex than previously thought and that the understanding of the mechanisms of toxicity requires proactive multidisciplinary research initiatives to address the impact of NP on human health. This requires the use of a cocktail of advanced analytical and bioanalytical techniques for a focused research studies directed towards the understanding of mechanisms of NP toxicity opening a new branch of analytical chemistry, namely analytical nanotoxicology. This paper intends to highlight how the analytical chemist can contribute in the exciting field of in vitro mechanistically-based nanotoxicology research by alternative non animal methods such as cell cultures. An example showing the role played by unique radioanalytical and advanced spectrochemical techniques (ICPMS and GFAAS) in in vitro nanotoxicology research is presented. It concerns biokinetics aspects of metallic cobalt NP and cobalt ions in mouse fibroblast Balb/3T3 cell line, an in vitro model used to assess the cytotoxicity and carcinogenic potential of chemicals, in relation to the induced cytotoxic effects.

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OL-11 SPECTROCHEMICAL INVESTIGATION OF ELECTRODE COATINGS DESIGNED FOR ELECTROCHEMICAL WASTE WATER TREATMENT

János Kristóf1, Erzsébet Horváth2, Lourdes Vázquez-Gómez3, Achille De Battisti3 1University of Veszprém, Department of Analytical Chemistry, H-8201 Veszprém, P.O.Box 158, Hungary 2University of Veszprém, Department of Environmental Engineering and Chemical Technology, H-8201 Veszprém, P.O.Box 158, Hungary 3University of Ferrara, Department of Chemistry, I-44100 Ferrara, Italy

Conductive metal oxide film electrodes as dimensionally stable anodes (DSA®) have been of high technological importance since the sixties. The class of thermally prepared oxide electrodes on titanium metal support contain a catalytically active noble metal oxide (IrO2 or RuO2) stabilized by TiO2, ZrO2, Ta2O5 or SnO2. The most important commercial types are the RuO2/TiO2 and the IrO2/Ta2O5 systems used as electrodes in the chlor-alkali industry and in other industrial processes where oxygen evolution is one of the electrode reactions at the anode. Recent research has been carried out on the feasibility of electrochemical incineration of organic pollutants in industrial waters of high COD and TOC contents (e.g. due to saccharides, polyphenols, tannins in the food industry) and of strongly polluting chemicals (e.g. chloro- and nitrophenols in synthesis waters). A carefully planned electrochemical treatment can be a realistic alternative to biological attacks on organic pollutants of biorefractory nature. In the present work the formation mechanism of the individual film components and that of the most important mixed oxide systems suitable for water purification applications and in electrometallurgy is studied under in situ conditions using thermogravimetry combined with mass spectrometry (TG-MS), diffuse reflectance Fourier transform infrared spectrometry (DRIFT), and emission infrared spectroscopy (IRES). With the mass spectrometric ion intensity curves the changes in the composition of the gas phase can be followed. Information on the evolution of the solid phase can be obtained with the infrared spectroscopic techniques. The sol-gel method of film preparation used has several advantages over other techniques such as low-cost processing, easy control of film thickness, and its flexibility when the kinetics of phase formation has to be controlled, or third components has to be added. Electrochemical and surface properties of thermally prepared oxide anodes depend strongly on the conditions of preparation (e.g. substrate pretreatment, precursor solution composition, and firing temperature). Cracking/combustion of retained solvent, formation and decomposition of surface carbonyls, carboxylates, intramolecular hydrolytic reactions, combustion of trapped carbon are among the processes influencing surface morphology and electrochemical performance of the final coating. Knowing the history of film evolution, preparation conditions can be optimized for superior performance.

[1] E. Horváth, J. Kristóf, H. Nasser, R.L. Frost, A. De Battisti, Á. Rédey: Applied Surface Science, 242 (2005), 13-20. [2] J. Kristóf, T. Szilágyi, E. Horváth, A. De Battisti, R.L. Frost, Á. Rédey: Thermochimica Acta, 413 (2004), 93-99. [3] J. Kristóf, A. De Battisti G. Keresztury,E. Horváth, T. Szilágyi: Langmuir, 17 (2001), 1637-1640.

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OL-12 ANALYTICAL CAPABILITIES OF ETV-ICP-OES METHOD APPLIED FOR ANALYSIS OF CERAMIC, BIOLOGICAL AND ENVIRONMENTAL MATERIALS Jürgen Hassler1, Peter Barth1, Gyula Zaray2,3, Karol Florian4 1ESK Ceramics GmbH & Co KG, Kempten, Max Schaidhauf Str. 25, D-87471, Kempten, Germany 2Department of Inorganic and Analytical Chemistry,. Eötvös University, P.O. Box 32, H- 1518 Budapest, Hungary 3Joint Research Group of Environmental Chemistry of Hungarian Academy of Sciences & Eötvös University, P. O. Box 32, H-1518 Budapest, Hungary 4Department of Chemistry, Faculty of Metallurgy, Technical University of Kosice, Letna 9, SK-042 00 Kosice, Slovakia

The electrothermal vaporisation-inductively coupled plasma-optical emission spectrometry (ETV-ICP-OES) is a widely used analytical system for elemental analysis of solutions and solid samples. The ETV-units developed for atomic emission spectrometry as sample introduction system can be divided into two groups. The vaporizer is located directly below the ICP-torch or a transport tube with length of about 0.4 – 0.8 mm is used to connect the vaporization and the excitation units. However, in the second case it is necessary to form a well transportable aerosol. For this propose the hot vapours are mixed with cool argon gas. This type of ETV-unit was developed by Spectral System (Fürstenfeldbruck, Germany).

This modern ETV-ICP technique is a powerful and cost-effective tool for routine analysis of solid samples. The range of applications is extremely wide, especially if modifiers are also used. Ceramic raw materials and sintered products, sediments, geological and environmental samples as well as biological materials such as human, animal and plant tissues, blood, serum and all kinds of liquids or even oil were analyzed in different industrial and research laboratories.

In this paper the statistical evaluation of the routine determination of metallic impurities in boron nitride powder over a two year period will be discussed, representing 318 analytical runs with the same number of independent calibration functions. On the other hand the versatility of ETV-ICP-OES will be demonstrated via common calibration functions for different materials like graphite, silicon carbide, sludges, apple leaves, green alga and dried liquid standards. Zaray, Gy.; Leis, F.; Kantor, T.; Hassler, J.; Tölg, G.: Analysis of silicon carbide powder by ETV-ICP-AES. Fresenius J Anal Chem 1993, 346:1042-1046 Hassler, J.; Detcheva, A.; Förster, O.; Perzl, P. R.; Florian, K.: Working with a modern ETV-device and an ICP-CID-spectrometer. Annali di Chimica 1999, 89:827-836

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OL-13 BIOMARKERS: AN EFFECTIVE TOOL TO ASSESS HUMAN EXPOSURE TO CHEMICALS IN EVERYDAY LIFE Marina Patriarca, Antonio Menditto Department of Food Safety and Veterinary Public Health, Istituto Superiore di Sanità - viale Regina Elena 299 - 00161 Rome, Italy E-mails: marina.patriarca@iss.it; antonio.menditto@iss.it Chemicals are present in the environment due to natural and human activities and people may come in contact with them in several everyday situations: at home, at work, with food, beverages or because of other lifestyle habits, such as cigarette smoking. Particular concern arises for long term exposure to low concentrations which may cause more subtle effects, which are more difficult to recognise, and may affect even future generations through subtle alterations of the reproductive system and genetic heritage. In order to develop effective measures for both the protection of human health and the promotion of sustainable development of trade and industry, it is necessary to acquire reliable information on the exposure of the general population, with particular attention to more susceptible groups. Whereas environmental and food monitoring allows to identify the presence of contaminants, programs of biological monitoring based on the determination of suitable biomarkers in biological fluids, tissue and excreta provide direct information on the internal dose, the severity of effects and, in some cases, the individual susceptibility to a health threat from a certain substance. However, despite its advantages, several difficulties hinder a wider application of biological monitoring for risk assessment, including lack of specific biomarkers, the complexity of the process required for their validation and ethic constraints. Some aspects of the challenges related to the development and application of biomarkers to the assessment of human exposure to chemicals will be reviewed and discussed.

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OL-14 EMERGING CONTAMINANTS OF WATER RESOURCES: PESTICIDE METABOLITES AND PHARMACEUTICALS Paola Bottoni Istituto Superiore di Sanità, Viale Regina Elena 299, 00161 Roma, Italy

The widespread and growing use of pesticides in agriculture over the last forty years has caused the deterioration of surface and groundwater bodies with adverse impacts on aquatic and terrestrial ecosystems (death of fish, crustaceans, birds, imbalances and alterations of food webs) and with worsening of drinking water quality in many countries. The increasing awareness of the risks posed by pesticides has by far prompted both scientific research, aimed mainly at defining levels of contamination, environmental behaviour, biological effects and hazards, and regulatory activities intended to limit emission in water bodies, to safeguard the health of aquatic organisms and man and the availability of safe drinking water. More recently, the importance of degradation products of pesticides as water contaminants has been highlighted. When released into the environment, pesticides may be transformed by abiotic and biotic degradation processes into plenty of unknown intermediate compounds, called as a whole “metabolites”, which generally undergo mineralization. In some cases, it has been demonstrated the formation of persistent, mobile and toxic metabolites which may reach water bodies with common transport patterns, as surface runoff, leaching and aerial spray drift. Thus, pesticide metabolites may pose a risk if water is used for drinking. So far, experimental data published in the literature and monitoring activities indicate that metabolites may be detected in water bodies at frequencies and levels higher than their parent compounds, often at concentrations of tens of µg/L along with and, in some cases, in the absence of their corresponding parent compounds. The importance of metabolites as water pollutants has been definitively established since 1998, when they were included in the “pesticide” parameter of the European Directive 98/83/CE for drinking water protection. According to this Directive, “relevant” metabolites undergo the same MACs of 0.1 and 0.5 µg/L of pesticides (Annex I, Part B, note 6). Even so, metabolites are rarely included in routine monitoring plans both for the large numbers of pesticides that each country has yet to take into account and for the difficulty to identify the relevant ones, as the key intrinsic parameters and the ecotoxicological and toxicological data which are required to define their environmental behaviour and their biological significance are rarely available in the open literature. A possible approach aimed at the selection of metabolites of environmental and health concern is presented. Furthermore, some data on the presence of these compounds in Italian surface and ground waters are shown. Another emerging category of water contaminants is represented by human and veterinary pharmaceuticals as well as by their bioactive metabolites. Similarly to pesticides, these compounds have a large and increasing use worldwide, but, on the other hand, they can reach water courses primarily through point sources of contamination as urban and hospital sewage systems, municipal sewage treatment plants, industrial discharges, zootechnical and aquaculture waste water, inadequate disposal of unused and expired products etc. Some studies indicate that common treatment technologies fail to remove several classes of pharmaceuticals, mainly due to the inactivation of biological systems and/or low adsorption of polar compounds to sewage sludge. As in the case of pesticides, pharmaceuticals include hundreds of heterogeneous molecules with very different chemical-physical properties and

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chemodynamic properties, such as soil and hydrolysis half-lives and soil partition constants (Koc and Kd) which are largely unknown. Hence, the possibility to define their environmental behaviour is mostly unfeasible. Some pharmaceuticals are known for their persistence in soil and water or for their resistance to biodegradation in sewage treatment plants (X-ray contrast compounds, cyclophosphamide, clofibric acid, ivermectin oxolinic acid, flumequine, oxytetracicline). Also pharmaceuticals have a wide spectrum of important biological properties, which, as a rule, are conceived as a benefit in terms of protection of human and animal organisms; so far, they have not been considered for possible side effects on environmental species, but only recently. It is known that pharmaceuticals may cause adverse effects on aquatic biota, as the development of antibiotic resistance in pathogenic microorganisms, the appearance of morphological alterations and sex impairment on aquatic organisms and fish due to antibiotics, hormones and contraceptives, respectively. Nevertheless, data sets on acute and chronic ecotoxicity show many gaps. The presence of some pharmaceuticals in water bodies has been frequently ascertained in these last twenty years. Generally, antibiotics, analgesics, antidepressants, hormones, estrogens, haematic lipid regulators, disinfectants, anti-inflammatory and anti-epileptic drugs etc. have been found at levels of ng-µg/L, both in surface water and groundwater and up to hundreds of mg/kg in sediments. Yet, the significance of these findings has not been adequately highlighted. Indeed, studies on environmental risk assessment are in progress and, in spite of the paucity of data, it has been concluded that some compounds may pose a risk to aquatic fauna (ibuprofen, acetylsalicilic acid, mefenamic acid, aminotryptiline, amoxicillin, dextropropoxyphen, fluoxietin, oxytetraciclin, paracetamol, propanolol and tioridazine). As regards indirect exposure through drinking water, a comparison of levels of pharmaceuticals in drinking water and therapeutic doses does not point to an actual risk for human health.

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OL-15 INFLUENCE ON ELEMENTAL STATUS BY HIP-ENDOPROSTHESES Michaela Zeiner1, Ilse Steffan1, Felix Lintner2, Wolfgang Schwägerl3, Peter Zenz3

1Department of Analytical Chemistry and Food Chemistry, University of Vienna, Währinger-straße 38, A-1090 Vienna 2Pathologisch-Bakteriologisches Institut, Sozialmedizinisches Zentrum, Otto Wagner Spital Baumgartner Höhe 1, A-1145 Vienna 3Orthopädische Abteilung, Sozialmedizinisches Zentrum, Otto Wagner Spital Baumgartner Höhe 1, A-1145 Vienna For many clinical purposes in modern medicine various devices or materials are applied, which contact tissue, blood and other body fluids, such as the synovial fluid. Adverse reactions between foreign or prosthetic surfaces and body components, such as blood and various types of tissues are limiting the application of biomaterials for medical purposes. Of the many materials applied for surgical implants the most common are metals and alloys in use for orthopaedic joint replacements, especially the total hip replacement. Cobalt-chromium-molybdenum alloys are still the most frequently used metallic components for joint replacements. In these cases considerable amounts of cobalt-chromium-molybdenum wear particles are released from the prostheses and they have to be regarded as a cause of long-time problems for the patient. The released wear particles are dissolved in the body fluids of the surrounding area. Via blood the metals mainly bound to transporting proteins are distributed in the whole body. Elevated metal concentrations have been found in blood (serum and plasma) as well as in urine. Partly the metals are excreted via urine, but to some extend they accumulate in the different organs. Till now there are only a few studies published concerning metal concentrations in human organs. Therefore our objective was to optimize the determination of seven relevant trace elements, namely Al, Co, Cr, Mo, Nb, Ni, and Ti in six kinds of organs (brain, heart, kidney, liver, lung, muscle) starting with drawing of the sample, sample work up and finally analysis by means of atomic spectrometry. The optimized method was then applied to determine the metal contents in organs of persons with various kinds of total hip replacements. The statistically significant differences obtained in this pilot study require the necessity for further studies in this field.

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OL-16 STUDY OF INTERACTION BETWEEN PB AND PHARMACEUTICALS BY CHELATING SORBENTS AND GFAAS

Katalin Zih-Perényi, Alexandra Lásztity L. Eötvös University, H-1518 Budapest 112, P.O. Box 32., E-mail: perkat@freestart.hu

Lead is common trace in pharmaceuticals. Beside the classical methods based on coloured precipitation reaction, the European Pharmacopoeia prescribes the atomic absorption spectrometric tests for some drugs and drug materials.

Lead can be retained on cellulose containing imino diacetic acid ethyl groups (IDAEC) by chelating processes. The rate of sorption is influenced by the concomitant chelating groups, which are derived from the drug matrix components or from its organic/inorganic contaminants. Therefore the sorption gives information about the existence of Pb-interaction with materials present in the sample. Supposing homogenous equilibrium, the distribution of lead between the different species of chelating agents, e.g. chelating cellulose and matrix, can be calculated. For this calculation the amount and the appropriate stability constants of the included ligands is necessary.

The stabilitity of different Pb-IDAEC species was determined by means of potenciometric titration curves. These curves graph the pH of the heterogeneous system in function of amount of NaOH added, representing the amount of protons evolved during interaction. Due to the higher affinity of lead toward imino diacetic acid (ethyl) groups in acidic range than expected, a protonated Pb-H-IDAEC form was supposed. In fact, lead is susceptible to form protonated and hydroxo-complexes with some ligands like imino diacetic acid (IDA) or ethylene diamin tetraacetic acid (EDTA).

A semi-on-line flow injection preconcentration was developed for lead including a minicolumn filled with IDAEC. The lead content, eluated by HNO3, was collected and determined by graphite furnace atomic absorption spectrometry (GFAAS). The detection limit of the method is 0.11 µgL-1 applying a preconcentration factor of 20.

The lead sorption of IDAEC measured in flow system can be predicted by values calculated for homogenous system and equilibrium. The measured and evaluated retention at different pH-s were in accordance. The effect of drug matrix, like penicilamine, acethyl-cystein, citrate, Na-alendronate, ampicillin, Captopryl, etc. was investigated. The variables were the type of pharmaceutical, the concentration of drug solution and the pH. The calculation was performed by the stability constant of the drug, or, if it was not available, the stability constant of low MW molecule with similar functional groups was applied. The theoretical sorption values provided by the model, exhibit good agreement with experimented ones.

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TUESDAY, 25 OCTOBER 2005

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OL-17 THE MEASUREMENT OF CHROMIUM SPECIATION IN ENVIRONMENTAL POLUTION AND ITS IMPACT UPON HUMAN HEALTH. Les Ebdon University of Luton, Park Square, Luton, LU1 3JU; E-mail: Les.Ebdon@luton.ac.uk Exposure to chromium has long been association with increased incidence of cancer. In particular industries, excess lung cancer deaths and cancers of the respiratory system have been associated with exposure to chromium 6. As a consequence, hexavalant chromium compounds are heavily regulated through the European Union Dangerous Substances Directive. By contrast, chromium 3 is regarded as being essential for health and has long been associated with the, so called, glucose tolerant factor, said to be absent in diabetics. There is therefore a clear need for rapid and simple methodolgy to distinguish between the different species of chromium in a variety of environmental samples. This paper will describe the newly developed sample introduction method for the speciation of chromium using quadrupole-ICP-MS. The modified sample introduction system consists of a microbore column between the solvent delivery system and the ICP. Given the necessarily low operating sample flow rates of the column, a microconcentric nebulizer (MCN) and, preferably, a direct injection nebulizer (DIN) were used as the interface between the column and the ICP-mass spectrometer. These capillaries were extensively tested in terms of their matrix removal, pre-concentration and chromatographic behaviour. The effects of sample pH, gas flow rates and matrix influences are reported. Fused silica capillaries were treated with three different anion- and cation exchange reagents: 3-aminopropyltrimethoxy silane (AP-TMS), 2-(4-chlorosulfonylphenyl)-ethyltrichlorsilane and 2-acrylamido-2-methylpropane sulfonic acid. The latter two proved to be the most efficient reagents for the separation of Cr(III) and Cr(VI). Cobalt was used as internal standard and its signal intensity was continuously monitored and used to correct for signal drift and instrumental instability. Excellent results were obtained in terms of repeatability. The precision (coefficient of variation) obtained for different amounts of Cr(III) and Cr(VI) was between 1.7 and 2.1% using the DIN and between 2.8 and 3.7% for the MCN, respectively. Detection limits for Cr(III) and Cr(VI) were 8 ng L-1 and 7 ng L-1 respectively. Accuracy was assessed by analysing a certified reference material welding dust CRM 545. The results obtained were in good agreement with those certified.

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OL-18 METHOD DEVELOPMENT FOR MERCURY SPECIATION STUDIES IN FISH SAMPLES INTENDED FOR HUMAN CONSUMPTION László Abrankó, Zsuzsa Jókai, Béla Kmellár, Péter Fodor

Department of Applied Chemistry, Corvinus University of Budapest, Villányi út 29-33, 1118 Budapest, Hungary; E-mail: laszlo.abranko@uni-corvinus.hu There is a growing interest in mercury speciation in samples of human food products due to varying toxicity of the different chemical forms of mercury. Organic mercury compounds, of which methylmercury is the most common, are of special concern because of their enhanced toxicity. The route of human exposure to methylmercury is mainly through the diet, especially via fish that bioaccumulate this compound. In response to growing demand, several analytical techniques have been developed over the past decade to speciate organomercurials in biological samples. In the present work, the application of a cost-efficient analytical method (SPME-GC-pyrolysis-AFS system) for the determination of MeHg in typical dietary seafood samples is described. The study focused on investigating the modifications necessary for well-established methods involving alkaline digestion in order to use aqueous phase phenylation and SPME sampling to analyze food samples with complex matrices containing MeHg in the range of 100 ng g-1.

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OL-19 SPECIATION ANALYSIS OF SELENIUM IN ALFALFA SPROUTS BY TWO-DIMENSIONAL HIGH-PERFORMANCE LIQUID CHROMATOGRAPHY WITH PARALLEL ICP-MS AND ESI-MS DETECTION Valeria Gergely1, María Montes-Bayón2, Rita Szalma1, Alfredo Sanz-Medel2, Péter Fodor1 1Corvinus University of Budapest- Department of Applied Chemistry, H-1118, Budapest, Villányi ut 29-33, Hungary 2 University of Oviedo, Department of Physical & Analytical Chemistry, C/ Julián Clavería 8, Oviedo, E-33006 Spain Selenium is an essential trace element for humans and animals. Since the work of Clark et al. involving the role of selenium in cancer chemoprevention, potential "Se-delivery systems” have been explored. Research involving selenium has become an increasingly popular area in the research community in the last twenty years. A lot is known about the health implications of selenium, but there are still numerous unexplored areas. One problem with selenium as a supplement is a narrow range between the selenium toxicity level and beneficial level. At the proper levels selenium has been shown necessary for proper immune system functioning. Eating a healthy and nutritionally balanced diet is an important step towards achieving body’s nutritional requirements. The type of selenium species is a primary determinant of its absorption and subsequent utilization, called bioavailability. Organic selenium species (selenomethionine, selenocysteine and methylselenocystein etc.) have been shown more bioavailability than inorganic species. The major forms of selenium in foods are probably the amino acids bound to proteins. Selenomethionine is presumably the prevalent form in diets from plant sources. [1,2] In the present study the distribution pattern of selenium in alfalfa sprout was determined by the means of two dimensional high performance liquid chromatography with ICP-MS and ESI-MS detection. The sample was leached with water and the aqueous extract was fractionated by preparative size-exclusion chromatography. Selenium was found in high- and also low-molecular mass fractions. The chromatographic purity of the fraction was verified by reversed-phase chromatography with inductively coupled mass spectrometric detection. The retention time of the liberated selenium compounds in HPLC–ICP MS matched that of selenomethionine (SeMet) and Se-methylselnocysteine (MeSeCys), and the masses of molecular and fragment ions detected by HPLC–ESI MS also suggested that the selenium compounds were SeMet and MeSeCys. Also the study revealed the existence of various unidentified Se species in the sprout one of them was identified by ESI-MS. [1] L.C. Clark et al., JAMA-J. Am. Med. Assoc. 276 (1996) 1957. [2] C. Ip, J. Nutr. 128 (1998) 1845.

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OL-20 COMPREHENSIVE SELENIUM SPECIATION STUDIES IN Se-ENRICHED CHIVES (ALLIUM SCHOENOPRASUM) BY HPLC- ICP-MS Emese Kápolna1, Monika Shah2, Joseph A. Caruso2, Péter Fodor1 1Corvinus University of Budapest- Department of Applied Chemistry, H-1118, Budapest, Villányi ut 29-33, Hungary 2Department of Chemistry, University of Cincinnati, P.O. Box 0172, Cincinnati, OH 45221-0172, USA Chives (Allium schoenoprasum) separately enriched with 10 µg ml-1 Se(IV), Se(VI) and SeMet were studied from the possible change in the species distribution point of view arising from the enrichments. The highest selenium accumulation up to 700 µg Se g-1 was observed in the case of the Se(VI)-enriched sample on the basis of total selenium measurements. Three liquid chromatographic techniques were applied. For the separation of selenium containing proteins size-exclusion chromatography (SEC-HPLC), for speciation studies reversed-phase ion-pairing chromatography (RP-IP-HPLC) and for the separation of enantiomers of three selenoamino acids a crown ether column was coupled on-line to an ICP-MS. Proteins were precipitated from the chives for the SEC analysis. For speciation studies two extractions (perchloric-acid-ethanol and enzymatic) were carried out on chive samples. In every sample the SEC studies showed a fairly intensive peak in the high molecular weight range (125 kDa – 1.355 kDa). The extraction efficiency proved to be very poor (~ 30%) when applying the perchloric-acid-ethanol extractants compared to the proteolytic enzyme (~ 70%). The highest contribution of organic species (MeSeCys and SeCys2) was present in the enzymatic extracts of SeMet-enriched sample. The chiral speciation revealed the presence of the L- enantiomeric forms of selenoamino acids in both extracts. L-MeSeCys was the major enantiomer found in the perchloric acid-ethanol extracts, while analyzing the enzymatic extracts, besides L-MeSeCys, L-SeMet was also detected.

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OL-21 ARSENIC SPECIATION IN SAMPLES FROM FRESHWATER ORIGIN

Richard Schaeffer1, Csilla Soeroes1, Péter Fodor1, Walter Goessler2, Kevin A. Francesconi2, Doris Kuehneltb 1Department of Applied Chemistry, Corvinus University of Budapest, Villányi út 29-33, 1118 Budapest, Hungary; Email: richard.schaffer@uni-corvinus.hu 2Institute of Chemistry, Analytical Chemistry, Karl-Franzens-University, Universitaetsplatz 1, 8010 Graz, Austria

It is well known that various arsenic compounds are accumulated in the marine ecosystem, especially in algae, bivalves, and fish. During the last decades a lot of arsenic speciation studies have been carried out, mostly focusing on the analysis of marine biota in which the accumulated arsenic level is high enough for speciation analysis. Until recently, the biota of freshwater environments has been considered to contain low arsenic concentrations. Therefore, only a few speciation studies on freshwater ecosystem have been carried out.

We investigated the arsenic accumulation and distribution of arsenic species in a range of biological samples of freshwater origin including algae, plants, mussels, frogs, and various fish species collected in the water reservoirs in Hungary.

In the first part of the study, a river water food chain has been sampled from Danube and the arsenic speciation of the collected samples was carried out in order to compare their arsenic speciation pattern with the marine environment. The results suggest that the total arsenic accumulation of freshwater algae and mussels is comparable to the marine one. The dominating arsenic species in freshwater algae were arsenosugars, while plants contained only inorganic arsenic, similar to the marine ecosystem. But for fish and mussels a different picture was obtained. While only trace concentrations of arsenobetaine were found in freshwater mussels, it is a common arsenical in marine mussels Moreover, some thio-arsenosugars were found in the freshwater mussel samples, recently also reported for marine mussels.

In the second part of our work the most widely consumed fish species – carps and catfish - were collected in southeast Hungary, which is well known for elevated arsenic level in groundwater. In the carps, inorganic arsenic, methylarsonic acid and dimethylarsinic acid were found. In the catfish samples, which were grown in thermal water pools (150-190 µg As/l), mostly arsenobetaine was found. The various analytical problems faced during the work – e.g.: extraction efficiency, column recovery and the separation of the newly identified arsenic species – will be discussed in the presentation.

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OL-22 ISOTOPE DILUTION ANALYSIS MASS SPECTROMETRY FOR THE ROUTINE MEASUREMENT OF BUTYLTIN COMPOUNDS IN MARINE ENVIRONMENTAL AND BIOLOGICAL SAMPLES

Márta Üveges1, Pablo Rodríguez-González2, J. Ignacio García Alonso2, Alfredo Sanz-Medel2, Péter Fodor1

1Corvinus University of Budapest, Department of Applied Chemistry, 29-33. Villányi út 1118 Budapest, Hungary 2Department of Physical and Analytical Chemistry, University of Oviedo, C/ Julián Clavería 8, 33006 Oviedo, Spain

The use organotin compounds, particularly tributyltin (TBT), as a toxic additive in antifouling paints has been responsible of their anthropogenic widespread introduction into the aquatic environment and in the food chain. TBT and its degradation products (di- and monobutyltin) persist in water and particularly in sediments (where they are preconcentrated), killing not only the sea life attached to the hulls of ships but also other non-target organisms. Due to the extremely high toxicity and the endocrine disrupting power of these tin compounds, harmful effects in many marine species have been reported worldwide. As a consequence, several countries have adopted legislative restrictions to regulate the usage of TBT in antifouling paints. In fact, the EU has recently included TBT and its degradation products in the list of priority pollutantsi. This environmental problem shows the urgent need for validated procedures in the speciation of organotin compounds in waters, sediment and biota from different environmental compartments and especially in shellfish-growing areas. However, their reliable quantification in environmental samples, particularly in solid matrices, is still difficult due to the extremely low levels in which these compounds occur in the environment and the use of different derivatisation, extraction and clean-up procedures which are necessary to preconcentrate and separate those compounds from interfering substances. Due to the scarce number of certified reference materials (CRM´s) for organotin in the field of speciation the highly qualified primary methods such as Isotope Dilution mass spectrometry (IDMS) turn into the best alternative to validate the analytical methodologies. Using IDMS it is possible to correct for the majority of systematic errors that occur in a speciation analysis such as low derivatisation yields, non-quantitative separation procedures, signal drift, quantification measurement errors, etc, providing highly accurate and precise determinations. The estuary of the river Eo (Northwest Spain) has been used for some years for oyster farming but at present no data exist in this area on the levels of butyltin compounds in water sediments or biota. The aim of this work will be the study of the contamination by butyltin compounds in this area by applying IDMS. For this purpose it was employed a commercially available spike solution containing a mixture of mono-, di-, and tributyltin enriched in 119Sn. The proposed methodology allows the fast, reliable and simultaneous determination of the three butyltin compounds in different types of samples providing not only a higher quality of the analytical results in comparison with more classical calibration strategies but also a drastic minimisation of the time required both in the sample preparation steps and in the analytical measurement.

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OL-23 CONTRIBUTION OF TRAFFIC-RELATED SOURCES TO AEROSOL COMPOSITION IN DOWNTOWN BUDAPEST

Imre Salma1, Tünde Labuda1, Willy Maenhaut2, Gyula Záray1

1 Eötvös University, H–1518 Budapest, P.O. Box 32, Hungary 2 Ghent University, Proeftuinstraat 86, B–9000 Ghent, Belgium Research studies performed systematically in springs since 1996 yielded data sets for particulate matter (PM) in PM10–2.0 and PM2.0 size fractions that can be used for investigating the changes in aerosol composition, and for identifying the major aerosol sources in downtown Budapest. It is the atmospheric concentrations for elements (Na – U), black carbon (BC), and PM mass that were derived by sampling methods and devices, and by analytical methods that remained the same over the years. The comparison of the data was performed by ratio of medians for the daily concentrations of aerosol constituents in 2002 to that in 1996. It is seen that the median PM10–2.0 mass decreased by 25%. The change is essentially due to lower level of the crustal component (road dust), which was responsible for about 50% of the PM mass in 2002. Median concentration of some typical anthropogenic elements Pb, As, Br, V, and S exhibited substantially larger decrease than the PM. At the same time, the median concentration of Sb, Cu, Zn, Cr, and Ba showed very considerable increase. In the PM2.0 size fraction, the median PM decreased by 12%. Median concentration of Pb, Br, As, Co, S, V, and Se decreased markedly, while medians for Sb, Cu, Ba, Na, Mg, Fe, Ca, and BC increased substantially. In 1996, the main source of particulate Pb and Br in both size fractions was undoubtedly leaded gasoline. Phasing out leaded gasoline in Hungary, which was completed on 1 April 1999, resulted in a drop of the concentrations in the downtown by a factor of 3–4 within some weeks, and the mean values have remained practically at the same level thereafter. In 2002, the correlation coefficients between Pb and Br were insignificant which means that their main sources were already distinct. Nevertheless, the mean crustal enrichment factors (EFs) for Pb in both size fractions were still large, which suggest that its important sources include other – expectedly anthropogenic – source types. The largest increase in both size fractions was observed for Sb and Cu. It is the mechanical wear of brake linings of vehicles that emits these elements into the atmosphere. A representative mean Cu/Sb concentration ratio of 4.6 ± 1.2 was obtained earlier for brake linings, while mean Cu/Sb ratio of 4.3 ± 0.8 for the PM10–2.0 size fraction was derived for downtown Budapest in 2002. Mean PM2.0/PM10–2.0 concentration ratios for Cu and Sb indicate that their main source was friction. Correlation between Cu and Sb in the PM10–2.0 size fraction was 0.99; and both of them were also correlated strongly with BC, Ca, Fe, and Ba. All this suggests that emissions from brake linings were the major source of Cu and Sb in 2002, especially in the PM10–2.0 size fraction. It is inferred from the large crustal EFs for Zn that it had important anthropogenic emission sources. Zinc occurs in tires of vehicles in an amount of several percents. Particulate Zn showed medium correlation with BC, Ca, Fe, Cu, Sb and Ba in both size fractions implying that the road traffic was a major source for atmospheric Zn in the PM10–2.0 size fraction. This work was supported by the Hungarian Scientific Research Fund (T043348), and by the Belgian Federal Science Policy Office (EV/02/11A).

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OL-24 A MICROANALYTICAL PROCEDURE FOR THE CHEMICAL CHARACTERISATION OF THE INORGANIC CONTENT OF ATMOSPHERIC PARTICULATE MATTER

S. Canepari1, E. Cardarelli1, M. Catrambone2, C. Perrino2, A. Pietrodangelo2, M. L. Astolfi1

1Department of Chemistry, University of Rome “La Sapienza”, P.le Aldo Moro 5, 00185 Rome, Italy 2C.N.R. Institute of Atmospheric Pollution, Via Salaria km. 29,300, C.P. 10, 00016 Monterotondo Stazione (Rome), Italy

Chemical characterization of fine airborne particulate matter (PM) has a key role both in understanding its adverse effects on human health, materials and the ecosystem, and in supporting the environmental policies at local and global scale. Moreover, it constitutes the starting point for any source apportionment of PM and for the comprehension of particulate pollution episodes. Particularly, the characterisation of inorganic ions can be of help in understanding secondary pollution events, whereas trace metals constitute a reliable tool for interpreting anthropogenic primary pollution events and for carrying out the source apportionment of atmospheric particles. A microanalytical procedure for the analysis of the inorganic content of atmospheric particulate matter is described in this paper. The procedure involves the analysis of non-volatile ions by ionic chromatography (IC) and of elements by both X-ray fluorescence (EPD-XRF) and optical inductively coupled plasma (ICP-OES) on a single membrane filter. Atmospheric 24-h PM10 samples are collected on a Teflon filter membrane at the flow rate of 2.3 m3/h. The first step of the procedure involves the elemental analysis by EPD-XRF. Then the loaded filter is extracted in an acetate buffer solution and the extract analysed for anions and cations by IC and for elements by ICP-OES. The residual is mineralised by acid digestion and then analysed by ICP-OES. The use of the three techniques on a single filter, besides yielding a complete picture of the inorganic content of the collected particles, allows the development of a reliable quality control scheme, able to check and quantify, sample-by-sample, systematic errors in the leaching procedure. The application of this 3-step method to a sampling campaign carried out in central Italy is also described. Comparison of the results yielded by two (IC and XRF, ICP and XRF) or by all three of the employed techniques in the determination of the same metals (e.g. S, Fe, Mg, Mn) is also reported.

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OL-25 IONIC COMPOSITION OF URBAN AND RURAL BACKGROUND AEROSOLS IN HUNGARY Rita Ocskay 1, Imre Salma 1, Willy Maenhaut 2, Gyula Záray 3 1Eötvös University, Department of Chemical Technology and Environmental Chemistry, Budapest, H-1518 Budapest, P.O. Box 32, Hungary 2Ghent University, Institute for Nuclear Sciences, Proeftuinstraat 86, B–9000 Ghent, Belgium 3Eötvös University, Department of Inorganic and Analytical Chemistry, H-1518 Budapest, P.O. Box 32, Hungary

Atmospheric particulate matter (PM) originates from a large number of natural and anthropogenic sources, so it is a complex mixture of different constituents. The knowledge of the chemical composition of aerosols as a function of size is necessary to determine their effects on human health and climate, and to identify their major sources. The water-soluble fraction plays an important role in many atmospheric processes. It has an effect on both direct and indirect aerosol forcing of climate, and the ionic composition determines the acidity of particles which influences the rate of many chemical reactions in the liquid phase.

As part of a Hungarian-Flemish Intergovernmental research project on atmospheric aerosol two sampling campaigns took place in Hungary: (1) in downtown Budapest at a kerbside in spring 2002, and (2) at the continental (rural) background air monitoring station, K-puszta in summer 2003. During both campaigns Gent-type stacked filter unit (SFU) samplers were operated on a day and night basis to collect particles in separate coarse (2.0–10 µm) and fine (<2.0 µm) size fractions. One of the SFU samplers collected the size fractions on polycarbonate/polycarbonate filter pairs; and the other one on polycarbonate/Teflon filter sets. The SFU samplers were operated in parallel at both sites. The PM masses were measured gravimetrically; the filters from the former SFU sampler were analyzed by particle-induced X-ray emission spectrometry (PIXE) and instrumental neutron activation analysis (INAA) for up to 46 elements. Filters from the latter SFU sampler were analyzed by ion chromatography (IC) for major inorganic anions (MSA–, NO2

–, NO3–, Cl–, Br–, SO4

2–, oxalate) and cations (Na+, K+, NH4

+, Mg2+, Ca2+). The mean fine-to-coarse mass ratios were 1.41 and 0.58 for K-puszta and Budapest,

respectively. At the rural site the fine particles were mainly responsible for PM10 temporal variability, which was attributed to long range transport of air masses. In Budapest, vehicular traffic was the main source of both fine and coarse particles. At K-puszta, the water-soluble inorganic ions measured were responsible for 44% of the fine particulate mass, and the main ionic components were SO4

2– and NH4+ accounting for about 91% (62 and 29%, respectively)

of the fine ionic mass. In Budapest, the inorganic ions contributed to the total fine particulate mass with 26%; in addition to SO4

2– and NH4+ ions, NO3

– was also significant. The ions listed accounted for about 48, 22 and 21% of the fine ionic mass, respectively. In the coarse size fraction the contribution of the water-soluble ions was similar (16 and 19% at K-puszta and in Budapest, respectively), although the coarse particulate mass was three times larger at the kerbside than at the background site. The main inorganic ions were Ca2+, NO3

– and SO42–. By

comparing the corresponding concentrations obtained from PIXE/INAA analyses and IC, we determined the water-extractable part of the constituents. About half of the total amount of coarse Na and coarse Mg, and about 80% of coarse K existed in water-insoluble forms, while both fine and coarse Ca was practically completely water-extractable. Only about 80% of total fine and coarse SO4

2– was water-soluble suggesting that part of the sulphur was present in

43

other than water-soluble form. The molar ratios of NH4+ to SO4

2–, and to NO3– in the fine

fraction indicated complete neutralization to (NH4)2SO4 and NH4NO3. According to the cation-to-anion ratios the coarse particles were alkaline, while the fine particles were slightly acidic or neutral. The anion deficit observed for fine particles was likely caused by water-soluble organic acids, and for the coarse particles it was due to carbonates.

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OL-26

SPECIATION OF NITROGEN COMPOUNDS IN NANOSCOPIC FINE AEROSOL SAMPLES USING TXRF-NEXAFS János Osán1, S. Török1, B. Beckhoff2, G. Ulm2, C. Abete3, R. Fuoco4 1KFKI Atomic Energy Research Institute, P.O. Box 49, H-1525 Budapest, Hungary 2Physikalisch-Technische Bundesanstalt, Berlin, Germany 3CNR-ICCOM, Pisa, Italy 4Department of Chemistry and Industrial Chemistry, University of Pisa, Pisa, Italy Nitrogen compounds play a very important role in the atmosphere’s chemical conversion processes. While compounds in the gas phase can be measured by high temporal and spatial resolution using optical and remote sensing methods it is crucial to have analytical methods that enable to measure the aerosols from a short sampling period while retaining the information on the size distribution of the particles. A near edge X-ray absorption fine structure (NEXAFS) investigation in conjunction with total reflection X-ray fluorescence (TXRF) analysis of minute depositions on wafer surfaces is able to contribute to the speciation of the atmospherically important low-Z elements (C, N and O). TXRF using high flux and tunable synchrotron radiation for the specimen excitation is a powerful technique for the non-destructive analysis of nanoscopic (10–9 g) amounts of solid material collected on silicon wafer surfaces allowing sampling for short time (1–10 min) or sampling in areas with very low aerosol mass concentration. Aerosol samples were collected at different locations close to seashore in Italy and at the Italian base near Ross Sea (Antarctica). Particles were sampled on silicon wafers using a seven-stage May cascade impactor. Aerosol collection was performed only at stages 5, 6 and 7, with aerodynamic cut-off diameters of 1, 0.5 and 0.25 µm, respectively. The TXRF-NEXAFS measurements were carried out at the PGM monochromator beamline at the electron storage ring BESSY II. Based on linear combinations of standard reference spectra of (NH4)2SO4 and NaNO3, the ratio of ammonium and nitrate could be quantified. Nitrogen was present almost entirely as ammonium at stage 7. The ammonium to nitrate ratio shows a decreasing trend with increasing particle size in the range of 0.25–2 µm. More than 90% of nitrogen was present as nitrate in the continental stage 5 samples. The Antarctica samples collected at the same stage however showed a much lower nitrate to ammonium ratio (55% to 45%).

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OL-27 COMPARISON AND EVALUATION OF DIFFERENT SAMPLING HEADS FOR WELDING FUMES SAMPLING

Balázs Berlinger, Miklós Náray National Center for Public Health, Central Chemical Laboratory, H-1450 Budapest P.O.B. 22., E-mail: berlingerb@fjokk.hu

The health of workers in many industries is at risk through exposure by inhalation to toxic metals and metalloids. Nowadays among this health damaging factors great attention is devoted to toxic effects by exposure of welding fumes. It is reasonable, because at present, it is estimated that more than one million workers are employed as welders worldwide, with more than three million performing welding intermittently as part of their work duties. Industrial hygienist and public health professionals has to determine the effectiveness of measures taken to control workers’ exposure, and this is generally performed by making workplace air measurements. Sampling is the most determining part of this measurements, but choosing of the most adequate sampler is not always an easy task. Welding fume samples were collected in two Hungarian welding plants, where different steels are welded with manual metal arc (MMA) welding. Welding fumes were sampled with fix-point and with personal sampling technique with different sampling heads: IOM, CIS and cyclone of the Higgins-Dewell design. The IOM head is developed at the Institute of Occupational Medicine in Scotland, and it is a recommended international standard for personal inhalable particulate sampling. The conic inhalable sampling head (CIS) is also a widespread sampling head in Europe for sampling inhalable dust in workplace air. The HD cyclone is for collection of respirable dust. After the sampling the welding fume samples were digested with the mixture of nitric acid and hydrogen peroxide in a closed vessel microwave digestion system and the solutions were analysed with two different analytical methods. The Cr, Mn, Ni and Co content of the solutions were determined with inductively coupled atomic emission spectrometry (ICP-AES) and with inductively coupled plasma mass spectrometry (ICP-MS). Iron was analysed with only the ICP-AES method. Toxic metal concentrations of the workplace air were calculated for all kind of the samplers, and the results were compared of the two different inhalable sampling heads. Finally the aerosol particulate size distribution was determined between the inhalable and respirable fractions. The mean differences between the two inhalable heads were not greater than 15% for all of the measured metals. The Cr content was 60-80% in the respirable dust correlating to its amount in the inhalable fraction. This rate was 55-75% for Ni, 70-90% for Mn, 50-70% for Co and 45-75% for Fe.

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OL-28 INVESTIGATION OF THE CORRELATION BETWEEN POLLEN VIABILITY AND ITS ELEMENTAL COMPOSITION

Giancarlo Pepponi ITC-irst, via Sommarie 18, 38050 Povo, Trento, Italy

Different authors have shown that the atmospheric pollution reduces the viability of pollen [1,2]. Pollen has therefore been proposed as a bio-indicator. Viability can be measured by the tetrazolium (TTC) and the fluorochromatic reaction (FCR) tests to measure respectively the respiratory activity and the integrity of the cellular wall of the pollen grains. In previous studies the effects of pollution have been studied extensively, but not much information is available about the relation between the viability and the elemental composition of the pollen. The present study aims to ascertain the correlation between the elemental composition of such samples and the results obtained by the TTC and FCR colorimetric tests. For this purpose, pollens of two different taxa were collected in areas with contrasting levels of pollution. In the choice of the analytical techniques it was considered that the degeneration of the pollen might concern a subset of the sample and it is potentially caused by a great variety of polluting agents not known a priori. These chemical species can be essential elements, traces of heavy metals and substances of anthropogenic origin, which might be present in low concentrations. Moreover, such samples can easily be contaminated by aerosols. These issues indicate that for this study we require sensitive multielemental techniques with a high lateral resolution to characterize individual pollen grains. Total Reflection X–Ray Fluorescence Analysis (TXRF), Inductively Coupled Plasma-Optical Emission Spectroscopy (ICP-OES) and Mass Spectroscopy (ICP-MS) have been adopted for the elemental analysis of the sample after acidic digestion. TXRF and ICP-OES cover similar elemental and concentration ranges and could be directly compared. ICP-MS features lower detection limits than the two previously mentioned techniques and has given insight in the content of ultra traces. Time of Flight Secondary Ion Mass Spectroscopy (ToF-SIMS) and Synchrotron Radiation Micro X-Ray Fluorescence Analysis (SR-µXRF) have been used to gain distinct information from single pollen grains. ToF-SIMS offers the highest lateral resolution and surface sensitivity, but has a poor sensitivity for the heavier elements. SR-µXRF has adequate lateral resolution for the analysis of single pollen grains and exhibits higher sensitivity for heavier elements showing good complementarity to ToF-SIMS. Corylus avellana L. and Pinus nigra L. pollen has been collected in 6 different locations in the Province of Trento, Italy. Pinus nigra L. pollen has also been collected in the Province of Piacenza (Italy) following a protocol to ensure higher statistical representativeness. Multivariate statistical tools such as Principal Component Analysis (PCA) and Partial Least Squares (PLS) have been employed for data reduction and visualisation of the results. These methods also highlight outliers and emphasise the correlation between elements, sites, and were adopted to discover a correlation between the viability and the elemental concentrations. [1] J.H.B. Wolters and M.J.M. Martens, Effects of air pollutants on pollen, Bot. Rev., 53, 1987, 372-414 [2] O. Iannotti, G. Mincigrucci, E. Bricchi, G. Frenguelli, Pollen viability as a bio-indicator of air quality, Aerobiologia, 16, 2000, 361-365 [3] G. Pepponi, P. Lazzeri, N. Coghe, M. Bersani, E. Gottardini, F. Cristofolini, G. Clauser and A. Torboli, Total reflection X-ray fluorescence analysis of pollen as an indicator for atmospheric pollution, Spectrochimica Acta Part B: Atomic Spectroscopy, 59, 8, 2004, 1205-1209

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OL-29 EVALUATION OF GROUND LEVEL OZONE CONCENTRATION USING MULTIVARIATE METHODS A. Lengyel1,2*, K. Héberger3, L. Paksy1, O. Bánhidi1,4, R. Rajkó5

1Department of Analytical Chemistry, Institute of Chemistry, University of Miskolc H-3515 Miskolc-Egyetemváros, Hungary 2Co-operation Center of Material Sciences and Mechatronics, University of Miskolc, H-3515 Miskolc-Egyetemváros, Hungary 3Institute of Chemistry, Chemical Research Center, Hungarian Academy of Sciences, H-1025 Budapest, Pusztaszeri út 59/67, Hungary 4DAM STEEL Co., Miskolc, Vasgyári út 43, H-3540, Hungary 5Department of Unit Operations and Environmental Engineering, University of Szeged College Faculty of Food Engineering, P.O.Box 433, Szeged, H-6701, Hungary *Author to whom correspondence should be addressed. E-mail: akmla@gold.uni-miskolc.hu

Multivariate statistical methods including pattern recognition (Principal Component Analysis: PCA) and modeling (Multiple Linear Regression (MLR), Partial Least Squares (PLS) as well as Principal Component Regression (PCR) are usually applied for selection and classification measuring data. Some possibilities are shown at the field of evaluation of environmental data.

These methods were carried out to evaluate the state of ambient air in Miskolc (second largest city in Hungary). Samples were taken from near the ground at a place with an extremely heavy traffic. Two groups of measured data were investigated. In the 1st one the data were measured by a day, the 2nd one includes values of several pollutants and of meteorological parameters measured about during one month.

Although PCA is not able to determine the significance of variables, it can uncover their similarities and classify the cases. PCA revealed that it is worth to separate day and night data because different factors influence the ozone concentrations during all day. Of course, PCA can also choose the data at the similar weather condition days.

Ozone concentration was modeled by MLR and PCR with the same efficiency, if the conditions of meteorological parameters were not different (i.e. morning and afternoon). Without night data, PCR and PLS (suggest that the main process is not a photochemical but a chemical one. Keywords: principal component analysis (PCA), principal component regression (PCR), partial least squares (PLS), environmental data, ambient air, modeling of ozone concentration.

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OL-30 DRY AND WET DEPOSITION INTO THE LAKE BALATON*

Klára Polyák, József Hlavay† Department of Earth and Environmental Sciences, University of Veszprém, H-8201 Veszprém, P.O. Box 158. Hungary, E-mail: polyak@almos.vein.hu

Atmospheric aerosols have important in the biogeochemistry and transportation of

trace elements in the air. The atmosphere is an important vector of global metal transport between regions, from land to sea, and vice versa. Direct atmospheric deposition makes only a minor contribution to the total metal concentration of the lithosphere because of the large reservoir of these metals in soils and rocks. It is well known that environmental effects of aerosol particles depend on their particle size and chemical composition. The aerosol particle s influence the solar radiation transfer, could aerosol interaction, and control the optical, electrical and radioactive properties of the atmosphere. Hence, the measurement of these two parameters is of crucial importance.

Speciation of trace metals in aerosols is different from speciation in aqueous media. The deposition, transport and inhalation processes are controlled predominantly by the size of atmospheric aerosols.

The aim of this research was to apply a simply sequential leaching procedure to the distribution of elements in atmospheric aerosols collected on the membrane filters at the Lake Balaton. A three-stage sequential leaching was applied to establish the distribution of metals among (1) environmentally mobile, (2) bound to carbonate and oxides, and (3) bound to silicates and organic matter (environmentally immobile) fractions. The aerosol samples were collected at Tihany, Siófok and Keszthely. At Tihany were collected the samples with cascade impactor to determine the velocity of deposition. From velocity of deposition and metal content of each fraction was determined the dry deposition at the Lake Balaton. At Tihany we were to collect the precipitation with wet-only sampler.

For comparison with earlier findings, mobile fraction, and total elemental composition of aerosols collected at different places in Hungary (K-puszta, Veszprém, Kabhegy, Budapest) and in different time at Lake Balaton.

*In memory of Prof. József Hlavay

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OL-31 STATISTICAL ANALYSIS OF AIR POLLUTION IN ROMANIA USING LEAF SAMPLES

Zita Margitai, Mihály Braun, Martine Leermakers

1Department of Inorganic and Analytical Chemistry, University of Debrecen, H-4032 Debrecen, Egyetem tér 1., Hungary 2Departement Analytische en Milieuchemie, Vrije Universitiet Brussel Pleinlaan 2, 1050 Brussel The element composition of the atmospheric aerosol and flying, depositing dust can be used to characterize the urban environment. The distribution of particulates varies within wide-range. Toxic elements occuring in urban air are usually sorbed or incorporated in the particulate matter. Due to their effect on human health, heavy metals are widely investigated compounds of dust. Air pollution may be detected by bioindicators. Depending on the degree of impact and meteorological, topographical factors, crown shape, density leaf size, crown density, dust accumulates on the leaves in various amounts. Dust accumulated on the surface of leaves can be collected and analysed easily. Some species of trees, as bioaccumulators, lend themselves for detecting define element and dust pollutions. Based on the chemical composition of the dust removed from the leaves, the pollution level of major geographical areas can be determined and biomonitoring system can be developed. The studied sample collected by this method is less well-defined as the sample resulted by standardized procedure. The purpose of the present research was to investigate the air pollution by measuring the element concentration of dust removed from the leaves. Results were statistically evaluated using Linear Discriminant Analyses (LDA) to characterise the type and significance of air pollutions. Samples were collected in August 2004 from 16 Transylvanian towns in Romania. The examined species of trees were as follows: Tilia×tomentosa, Tilia×cordata, Tilia×platyphyllos. Dust was removed by distilled water using ultrasonic bath and shaker maschine. Ten trees were investigated from each town and the sampled leaf area was10-12 dm2. To ensure accurate comparisons, both leaf area and dry weight measurements were taken. The samples were digested with nitric acid and hydrogene peroxide. The concentration of Al, Ca, Fe, K, Mg, Mn, S were measured by ICP-OES (Iris Advantage), Ba, Cr, Cu, Pb, Sr, V, Cd were analysed by ICP-MS (Thermo Finnigan Element 2). Discriminant Analyses was performed using SPPS program. The values of the discriminant functions characterize the certain groups (towns). The distance between the group averages (centroids) shows the differences in the composition of the dust originating from various places. In order to interpret the multi-variate (15 dimensions) space cluster-analyses (CA) was used. The distance between the centroids was calculated as quadratic Euclidean distance was used and the agglomerative algorithm was the Ward method. These methods make possible to divide the 16 towns into 2 main groups, which may be recognized in space and time, too. The pattern of pollution determined by remote sensing in Romania was similar to that we measured analysing the dust deposited on the leaves.

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OL-32 TILIA LEAVES AS BIOMONITORS OF AIR QUALITY IN EUROPE Mihály Braun1, Zita Margitai1, Martine Leermakers2 1Department of Inorganic and Analytical Chemistry, University of Debrecen, H-4032 Debrecen, Egyetem tér 1., Hungary 2Departement Analytische en Milieuchemie, Vrije Universitiet Brussel Pleinlaan 2, 1050 Brussel Bioaccumulators are able to collect and preserve heavy metals during of their lifetime. Air pollution of cities may be detected by bioindicator organisms. Our aim was to investigate the inorganic composition of urban dust deposited on the surface of Tilia species in large cities in Europe. Airborne dust deposited on the leaf surface of Tilia trees were collected from cities across Europe, and analysed for concentrations of Al, Ba, Ca, Cd, Cu, Fe, K, Mg, Pb, V, Zn (mg kg-1 d.w. dust). Samples were collected in May-August 2003 from 8 European cities (Antwerp, Brussels, Munich, Debrecen, Oradea, Cluj-Napoca, Baia-Mare, Varna). Tilia is one of the few tree species, which are widespread in Europe. The morphology, canopy structure and the dust preserving epicuticular wax on the leaf surface make the Tilia species an ideal object of biomonitoring. Leaves can be stored in refrigerators for long time without any damage or moulding. Dust was removed by distilled water using shaker machine and ultrasonic bath. The water content of suspension was removed by evaporation in microwave oven. After the determination of dry weight, dust samples were digested with nitric acid and hydrogene peroxide. Concentrations of inorganic compounds were measured by ICP-AES, flame (FAAS) and graphite furnace (GFAAS) atomic absorption spectroscopy. Our aim was to develop a biomonitoring system using this Tilia species. It was demonstrated that the leaves of Tilia×tomentosa and Tilia×platyphyllos occurred in same location accumulate different amount of dust. Small differences of leaf surface may cause differences in dust preserving ability. Only one of the species can be used to map of the heavy metal immission. Comparing the different Tilia species, the concentration of inorganic elements in the dust did not differ significantly. The composition of dust is independent of the species, thus all the three Tilia species can be used making the concentration maps. Classification of sampling sites was performed via a multivariate statistical procedure termed linear discriminant analysis (LDA) to characterise air inorganic quality characteristics and their relationships and the role of each element in differentiating between the cities. Our results suggest that elemental composition leaf-trapped dust reflect various environmental factors including local pollution sources (industry, traffic), city structure and climatic/meteorological features and can be used as a reliable indicator in air quality assessment and monitoring on local to regional spatial scales.

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OL-33 RIVERINE SEDIMENTS AS INDICATORS OF POLLUTION HISTORY István Papp1, Mihály Braun2

1Department of Mineralogy and Geology, University of Debrecen, H-4032 Debrecen, Egyetem tér 1, Hungary 2Department of Inorganic and Analytical Chemistry, University of Debrecen, H-4032 Debrecen, Egyetem tér 1, Hungary The mining and industrial activities in the Upper Tisza catchment area cause heavy metal pollution from time to time. The origin of the heavy metals (Cu, Pb, Zn) are the sulphide ores and the mining waste. These metals occur mostly in particulate forms in the river waters, such as sulphide grains or adsorbed on clay minerals, Fe- and Mn-oxyhydroxides. When the Tisza reaches the Hungarian Great Plain, its kinetic energy decreases and the suspended matter settles. The effects of pollution are stronger near the Hungarian-Romanian border, because most of the particulate material settles here. The pollutants in the river water are diluted downstream, so its threat to the environment is less in the Middle and Lower Tisza area. The heavy metal-bearing materials buried into the sediment, therefore historical data of pollution can be obtained by stratigraphic sediment analysis. The aim of this study was to estimate the reproducibility of the applied sampling method in a shallow oxbow lake. Our study area was the Boroszlókerti-Holt-Tisza, an oxbow lake on the floodplain of the upper part of the Tisza. This oxbow lake is flooded by the river every year. Sediment core samples were taken in each year by using a modified Livingston piston corer. Sub-samples were taken in 0,5-1 cm intervals. The sedimentation rate in the Tisza oxbow lakes is 2-3 cm·y-1, so the high-resolution subsampling method allows us to obtain historical data in year-scale. The samples were air-dried and digested with nitric acid and hydrogen peroxide. The concentrations of 20 elements were determined by ICP-OES and FAAS methods. The analytical results were evaluated by principal component analysis. High levels of anthropogenic metals indicate pollution events that were preserved in the sediment profile. The concentration peaks downcore can be well identified in all sediment samples, so the pollution history can be recorded from the sediments even in the case of shallow lakes. The results show that severe toxic heavy metal pollution occurred in the past, which were even more serious than Baia Borşa mine disaster in March 10, 2000. The applied multivariate statistical methods also allowed to distinguish between the alluvial and lake sediment layers in the core profile, and to obtain additional biogeochemical data, concerning the correlation of the elements.

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OL-34 DETERMINATION OF RARE EARTH ELEMENTS IN HUNGARIAN WINE SAMPLES AND BENTONITES USED FOR THEIR PURIFICATION BY DOUBLE FOCUSING SECTOR FIELD INDUCTIVELY COUPLED PLASMA MASS SPECTROMETRY Victor G. Mihucz1,2, Enikő Tatár2,3, István Virág3, László Rácz4, Gyula Záray1,2,3 1Joint Research Group of Environmental Chemistry of Hungarian Academy of Sciences & L. Eötvös University, P. O. Box 32, H-1518 Budapest, Hungary 2Hungarian Satellite Centre of Trace Elements Institute to UNESCO, P. O. Box 32, H-1518 Budapest, Hungary 3L. Eötvös University, Department of Inorganic and Analytical Chemistry, P.O. Box 32, H-1518 Budapest, Hungary 4 Károly Eszterházy College, Department of Chemistry, H-3300 Eger, Leányka u. 1. Wine is of a great economic importance, however, falsification, wrong declaration and blending of wines cause a serious financial shortage. Therefore, there is a strong need to develop fingerprint techniques assisted by mathematical statistical methods in order to determine the provenance of wines. Rare earth elements (REEs) would be ideal candidates for fingerprinting as they have similar chemical properties. The REE concentrations of four Hungarian white and red young wine samples collected from the viticulture area of Eger, Hungary, as well as of four bentonite samples (Gelbenton, Evergel, BW200 and Tükrös) were determined by double focusing sector field inductively coupled plasma mass spectrometry. Special attention was paid to the selection of the bentonite samples: Gelbenton is a fibrous purified bentonite, Evergel contains additives like egg albumin, caseine. BW200 and Tükrös are Na bentonites powders originating from Germany and Hungary, respectively. Wine and bentonite samples underwent to microwave-assisted digestion and fusion by lithium metaborate prior to the determination of their REE contents, respectively. The samples were analysed by applying internal standardisation with In in low-resolution mode (R = 300). The red wine samples were generally more concentrated for the REEs than the white ones. The effect of clarification on the REE concentrations was investigated by extracting the wine samples with the four above-mentioned, commercially available, bentonite samples. The simulated wine purification process consisted of adding adequate amounts of bentonites to the samples followed by stirring and a decantation period of 5 days, as it is a usual practice in oenology. For the evaluation of the effect chondrite normalisation was applied for the REE results. On the basis of the results, the Gelbenton and Evergel samples are recommended for purification as they did not alter much the REE concentrations of the purified wine samples, thus REE may be used as fingerprints for the determination of wine provenance by chemometric evaluation.

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WEDNESDAY, 26 OCTOBER 2005

54

OL-35 SEPARATION OF CHROMIUM(VI) USING COMPLEXATION AND ITS DETERMINATION WITH GFAAS IN ENVIRONMENTAL SAMPLES József Posta1, Áron Béni1, Roland Karosi1, Vasil Andruch2, József Balogh3

1Department of Inorganic and Analytical Chemistry, University of Debrecen, Egyetem tér 1, H-4032, Debrecen, Hungary. E-mail: postaj@tigris.unideb.hu. 2Institute of Chemistry, P. J. Šafárik University, Moyzesova 14, SK-04154, Košice, Slovak Republic. E-mail: andruch@upjs.sk 3Department of Chemistry, College of Nyíregyháza, Sóstói u. 31/b, H-4400, Nyíregyháza, Hungary. E-mail: baloghj@zeus.nyf.hu

In acidic medium and in the presence of chloride ions 2-[2-(4-methoxy-phenylamino)-vinyl]-1,3,3-trimethyl-3H-indolium chloride basic dye forms complex with Cr(VI). The optimum conditions (pH, concentration of Cl- and the complex forming reagent) of the separation and extraction of Cr(VI) into toluene using this basic dye as a complexing reagent have been determined and the possible interferences of Ca, Mg, Na, K, Cr(III), Ni, Pb, Hg, Mn, Al, Cu have been studied. An electrothermal atomic absorption spectrometer (GFAAS) was used for the determination of Cr(VI). The detection limit of the method for Cr(VI) found to be 0.15 µg dm-3 and RSD for spiked drinking water was better than 3%. Diperoxo chromium oxide is produced by reaction of hydrogen peroxide on chromium(VI). Diperoxo chromium oxide creates a complex with ethylacetate, while chromium(III) remains in an unchanged form in the aqueous phase. By this means chromium(VI) can be extracted from the aqueous phase. The optimal conditions of Cr(III)-Cr(VI) separation, as well as the chromium content of the ethylacetate phase were determined with graphite furnace atomic absorption spectrometry. In the second extraction of Cr(VI) from ethylacetate back into a water phase an additional preconcentration of chromium(VI) can be carried out. Because of the matrix free content of ethylacetate after the first extraction, with this separation a really extensive preconcentration of chromium(VI) can be realized.

An extraction followed by spectrophotometric method was successfully applied on a new extraction-atomic spectrometric method for the separation and determination of hexavalent chromium using GFAAS method. The suggested method improved the original spectrophotometric method’s detection limit at least by 3 orders of magnitude (50 ng L-1). With the twofold extraction and with the increasing of sample volumes a significant preconcentration of hexavalent chromium can be reached, wherewith the detection limit can be improved furthermore.

Developed analytical methods were successfully applied to determine chromium(VI) in rainwater, drinking water, sea water, waste water and soil extracts.

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OL-36 SPECIATION ANALYSES OF VANADIUM IN ETNEAN DRINKING WATER NETWORKS BY SPE AND ETAAS E. Veschetti, D. Maresca, L. Lucentini, E. Ferretti, G. Citti, M. Ottaviani Istituto Superiore Sanità, Dipartimento Ambiente e Connessa Prevenzione Primaria, Rome (Italy), Monitoring activities carried out since 1994 [1-2] showed the presence of significant levels of vanadium in drinking waters delivered in a lot of Etnean towns. The highest concentration (160-220 µg/L) was found in ground waters collected in the drainage gallery Ciapparazzo located on the Northwestern flank of Mt. Etna in Bronte area (Catania, Italy). This drainage gallery is an important water source for several towns of the Etnean province with a flow rate of near 500 L/s. Even though the toxicological assessment of vanadium is still in progress, experimental findings so far described in literature suggest that its presence in drinking waters may cause adverse effects when its concentration is much greater than a few tenths of micrograms per litre. Moreover, the toxicity of this element seems to increase as valance and solubility of its species increase [3-6]. On account of the different toxicological behaviour of V(IV) and V(V), which are the only possible oxidation states in aqueous mediums, a research project was set up to evaluate the ratio between their concentrations before and after disinfection treatments (chlorination or UV irradiation). Acquired data allowed evaluating the effect of residence time and disinfection treatment on possible species interconversion. In this article we discuss results obtained from the fractional determination of the two species in samples collected in the most representative sites of the drainage gallery and the distribution network. [1] L. Minelli, E. Veschetti, S. Giammanco, G. Mancini, M. Ottaviani; Microchem. J. 67 (2000) 83-90. [2] D.M. Previtera; Boll. Chim. Igien. 53 (2002) 309-326. [3] G.R. Willsky, D.A. White, B.C. McCabe; J. Biol. Chem. 259 (1984) 13273-13281. [4] B.Z. Zhong, Z.W. Gu, W.E. Wallace, W.Z. Whong, T. Ong. Mutation Res. 321 (1994) 35-42. [5] A. Léonard, G.B. Gerber; Mutation Res. 317 (1994) 81-88. [6] K. Kustin, D.L. Levine, G.C. McLeod, W.A. Curby; Biol. Bull. 150 (1976) 426-441.

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OL-37 APPLICATION OF THE SPECIATION ANALYSIS BASED ON SEQUENTIAL EXTRACTION IN THE WASTE RECYCLING TECHNOLOGIES

János Lakatos, Viktor Zs. Baranyai, Attila Lengyel University of Miskolc, Department of Chemistry, Miskolc-Egyetemváros, Hungary The economic issue of the recycling and the environmental risk caused by the toxic components of the waste is determined by the concentration and the speciation of the toxic elements. The sequential extraction technique can highlight the mobility of the component and provide very useful information for the design of the recycling technology.

The sequential extraction test of the sulphide ore waste deposits can provided information to the manner of the transformation of low solubility component to high solubility one as a results of oxidation. The comparison of the mobility of component of the red mud an the bauxite provided clear indication for the transformation of the minerals during the Bayern process. The minerals of iron and manganese were transformed to a less soluble form, however the minerals of chromium become more soluble. The solubility of the aluminium minerals also improved. Based on the difference of constituents mobility a separation method for the iron and aluminium was established.

The sequential extraction test using acidic and alkaline solutions was applied for the speciation of the Fe, Zn and Pb in the electric arc furnace (EAF) dusts. It was found that the system consist of different mobility iron zinc and lead fractions. Based on the results the ways of the EAF dust recycling, the possibility of the separation of the iron, zinc and lead can be demonstrated.

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OL-38 INVESTIGATION OF TRACE ELEMENT DISTRIBUTION IN UNPOLLUTED AND POLLTED SOIL BY SEQUENTIAL EXTRACTION PROCEDURE: PARTICLE SIZE DEPENDENCE Ágnes Hargitai -Tóth1, Zsuzsanna Csikós - Hartyáni2, József Balogh3 1, 3College of Nyíregyháza, 4400 Nyíregyháza, Hungary, tothag@nyf.hu, baloghj@nyf.hu 2University of Veszprém, 8200 Veszprém, Hungary, csikosne@almos.vein.hu

The soil is a complex heterodisperse system. The distribution of different soil components, such as minerals, organic and amorphous materials generally shows maximum or minimum in different particle size fractions. Thus it is expected that the distribution of trace elements bounded to different soil components is also dependent on the particle size distribution.

The species is a well defined atomic, ionic or molecular structure in the solid or liquid phase of the soil. Species can be determined among others operationally by sequential extraction procedure. The procedure starts with the mildest reagent to remove the most soluble fraction then a series of extractants of continuously increasing strength are applied to remove the more and more tightly held metals. Sequential extraction procedure divides the total metal content into fractions (species) of different mobility, and it shows the distribution among these fractions. The mobility of the trace element species is very important from the point of view of phytotoxicity and food chain. The mobile (or labile) metal species can be considered to be more active biologically than immobile fractions.

In this work sequential extraction procedure was performed for determination of distribution of trace element species (water soluble, exchangeable, weak acid extractable, reducible, oxidizable and residue) in different particle size fractions of a chernozem soil (unpolluted and polluted with Cd, Pb and Cr) covered with lime (Tiszalök). The soil sample was separated into the following particle size fractions: >5, 5-20, 20- 45, 45-80, 80-160, 160-315, 315-800, >800 µm. The mineral composition was characterised by X-ray diffractometry. The quantity of trace elements (Ni, Zn, Cu, Zr, Y, Ba, Sr, Cd, Pb, and Cr) in the solid phase was measured by X-ray fluorescence spectrometry. The concentrations of trace element species in the liquid phase were determined by AAS method. Our results show that the trace element species for the examined soil sample can be classified into two groups. Sr, Ba, Cr, Zr and Y (first group) were primarily present in the residual fraction of 20-45 µm particle size fraction. The greatest amounts of Ni, Zn, Cu, Pb and Cd (second group) were found in particle size fraction less than 5 µm in form of oxidizable (referring to organic bound trace elements) and reducible (regarding trace elements chemisorbed on the surface of Fe-Mn-oxides) species. The effect of contamination (Cd, Pb or Cr) on the distribution of the species of not polluting elements (Ni, Zn, Cu, Y, Zr, Ba, Sr) in the different sizes of the particles was insignificant. On the other hand the effect of contamination (Cd, Pb or Cr) on the distribution of the identical species (that is Cd, Pb or Cr) in the different sizes of the particles was very considerable. The pollution first of all increases the ratio of the mobile fractions. Through animal/human consumption of contaminated plants, the more mobile pollutants can enter easily the food chain. For this reason soil pollution is a source of danger to the health of mankind.

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OL-39 IODINE DETERMINATION IN DIETARY SUPPLEMENT PRODUCTS BY TXRF SPECTROMETRY Imre Varga Eötvös Loránd University, Institute of Chemistry, H-1518 Budapest P.O.Box 32, Hungary; vargaip@para.chem.elte.hu About two-thirds of the Hungarian population is living in areas where drinking water contains less than 0.025 mg/L I2. Iodine is an essential element required for the synthesis of thyroid hormones. The recommended daily allowance (RDA) is 0.150 mg for adults (WHO 1996). In case of iodine deficiency mineral water containing iodine and seaweed products could serve as the natural source of this element, among others. In the present study, the capability and limitations of total reflection X-ray fluorescence (TXRF) spectrometry for determination of iodine are reported, in the case of commercially available dietary supplements and differently processed seaweed (kelp) products. Multimineral tablets and kelp samples were digested using concentrated nitric acid before TXRF analysis. Extraction using a 25% ammonia solution was proved to be effective as an alternative sample preparation procedure for seaweed and dietary supplement tablets containing iodine. Precipitation of silver iodide, as a sample preparation step of mineral water, was applied to prevent loss of volatile iodine during solvent evaporation. Applying TXRF spectrometry iodine concentration was measured in iodine-enriched mineral water (94-100 mg/L), in seaweed samples and in tablets (26-5400 mg/kg).

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OL-40 THE MEASUREMENT OF RADIOACTIVITY IN DRINKING WATER

Maurizio Forte

ARPA Lombardia (Environmental Protection Agency) – Milano Department via Juvara 22, 20129 Milano, Italy; E-mail: m.forte@arpalombardia.it

Drinking waters generally contain variable amounts of radioactivity. Many radionuclides, like 40K and those belonging to the natural series of 238U, 235U and 232Th, are transferred to the water from the aquifer rocks by erosion and dissolution mechanisms.

In recent years, a great interest arose towards the natural radioactivity in drinking water. The European Council Directive 98/83/EC, subsequently enforced in Italian law, pushed public authorities to organize tap water surveys. Parameter values for radioativity levels in water are given, according to World Health Organization recommendations. Maximum concentration values for radon are separately proposed in a European Commission Recommendation.

For this reason, during the latest years, the Regional Environment Protection Agency (ARPA) has been performing an extensive monitoring of tap waters in the Lombardia district of Italy. Natural radioactivity content was measured to check the compliance with recent European and Italian rules.

Analyses were performed by an ultra low level scintillation counter equipped with an alpha-beta discrimination device. Specifically arranged procedures, requiring quick and easy pre-treatments, allowed to measure gross alpha and beta activity, 222Rn, 226Ra, and uranium isotopes concentrations in a relatively short time. Subsequently, more complex procedures for other natural radionuclides (210Pb, 210Po and 228Ra) were studied. Method performances were carefully tested by both internal validation procedures and international intercomparison exercises.

At first, tap water of the 13 largest towns was examined. Then, 30 sampling points in smaller towns were chosen to take into account the geographical and geological unhomogeneity of Lombardia which is partly a flat and partly a mountainous district. Obtained results showed that in most cases gross alpha activity was lower than the parameter value given by the cited regulations. When these values were exceeded, a more detailed survey was planned: at present three of them, in the western and south western part of Lombardia and in the city of Milan has been carried out while a third campaign, in its northern part, has just started.

Due to the widespread use of bottled mineral water, all major Italian brands have been examined as well, with special regards to those recommended for babies in lactation age.

Finally, the concern for terrorist actions, pushed our Institute to set up emergency procedures suitable to a rapid an reliable monitoring of public water resources.

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OL-41 THE CORRELATION BETWEEN URANIUM IN DRINKING WATER AND ITS EXPRESSION IN BIO-ASSAYS OF URINE, HAIR AND NAILS P. Marcus1,2, Z. Karpas2, O. Paz-Tal2, A. Lorber2, H. Sela2, Y. Hagag2 1Environmental Analytical Laboratory, Ben-Gurion University, Beer-Sheva 2Nuclear Research Center, Negev, Beer-Sheva, Israel L. Salonen, H. Komulainen, A. Auvinen, H. Saha, P. Kurttio, Finland Uranium is present in drinking water over a large range of concentrations depending on the origin of water source. The actual concentration varies from 10-2 µg/L for some springs to an extreme level of 104 µg/L for some private wells drilled in granite rocks. The limit of uranium has been set at 30 µg/L for communal water supplies by the US Environmental Protection Agency. Although only a small fraction of the ingested uranium (<1% for most of the population) is absorbed by the body, the total intake of uranium is reflected in bio-assays of urine, hair and nails. Urinalysis is routinely used to monitor the body content of uranium and bio-kinetic models have been developed for this purpose. Hair and nails are effective bio-concentrators and can thus be used to supplement or replace urine analysis. Inductively coupled plasma mass spectrometry (ICPMS) has proven to be a rapid, sensitive and accurate method for determination of the uranium content in these biological samples. The present study shows the correlation between the amount of uranium ingested in drinking water (the product of its concentration and the amount of water consumed) and its concentration in urine, hair and nails. The correlation between the 234U content in drinking water and its content in the bio-assays provides a direct link between the intake of uranium and its excretion from the body. This “built-in” tracer can be used for environmental and biological studies of uranium. The isotope ratio measurements were carried out by an MC-ICPMS located at GSI, Jerusalem thanks to the cooperation of Dr. Ludwik Halicz and his coworkers.

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OL-42 RAPID SEQUENTIAL METHODS FOR SEPARATION AND PRE-CONCENTRATION OF ANTHROPOGENIC RADIONUCLIDES FOLLOWED BY ALPHA-SPECTROMETRY AND ICP-SFMS DETECTION

Zsolt Varga, Zsolt Stefánka Hungarian Academy of Sciences, Chemical Research Center, Institute of Isotopes, H-1525, Budapest, P.O. Box. 77, Hungary, E-mail: varga@iki.kfki.hu and stefanka@iki.kfki.hu

The wide use of nuclear power created an increasing need for environmental monitoring of radionuclide pollutants. Actinides are considered the most toxic radionuclides in the environment, which can cause health hazards even in small concentration. Therefore radioanalytical laboratories throughout the world are frequently requested to measure actinides in matrices of environmental origin. Radiochemical methods, such as alpha spectrometry, have been most frequently used for determination of radionuclides. However, inductively coupled plasma mass spectrometry (ICP-MS) has become a powerful alternative technique because it combines the possibilities of multielemental and isotopic analysis with high sensitivity over a wide dynamic range and provides considerably shorter analysis time than alpha spectrometry. In our study we used for the determination of anthropogenic radionuclides both alpha-spectrometry (238Pu, 239,240Pu, 241Am) and ICP-SFMS (239Pu, 240Pu, 241Am, 236U). In order to reduce analysis time and consumption of chemicals a rapid and reliable sequential separation method has been developed. The method involves co-precipitation pre-concentration step and application of tandem extraction chromatography (UTEVA and TRU resins) separations. The method was employed for determination of actinides in different type of environmental samples e.g. natural water, soil and sediment. The analytical figures of merit of alpha spectrometry and ICP-MS analysis are compared and advantages and disadvantages of both methods are critically discussed.

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OL-43 ARCHAEOLOGICAL CHEMISTRY

Maria Perla Colombini Dipartimento di Chimica e Chimica Industriale, Università degli Studi di Pisa, via Risorgimento 35, 56126 Pisa, Italia, perla@dcci.unipi.it Since the mid Palaeolithic period, a wide variety of natural organic materials have been used as adhesives, sealants, painting and coating materials. Oils, natural resins and resinous materials played a prominent role, since their intrinsic properties meant that they could be used not only as adhesives, hydro-repellents, coating and sealing agents, but also as flavors, incense, ingredients for cosmetics, medicines and mummification balms. For their various applications, natural resins were used in their native form or, in some cases, subjected to hard-heating treatments and to distillation-type processes, in order to obtain denser and stickier materials, referred to as tars or pitches. Such heating treatments applied to natural resins and resinous wood deeply modify the chemical composition of the original material, inducing aromatisation, demethylation and decarboxilation reactions of the terpenoid compounds, with formation of new species, generally having lower chemical weight. For instance, one of the many applications of resinous material in Roman times was to treat the internal surface of amphorae in order to acquire waterproofing properties. The chemical characterization of such organic materials when properly integrated with related information from historical sources and archaeological data, has in the last few years considerably improved our knowledge of the crafts and technologies of the past, and has provided archaeologists with vital information. Moreover, if the chemical composition of objects in museums is known, this helps to assess the state of conservation of such objects along with any potential risk of degradation. The chemical analysis of organic substances in archaeological objects is generally based on the principle of “chemotaxis”, that is, on the presence of one or more specific (molecular biomarkers which have survived ageing or which are formed over the centuries as stable products of ageing processes. Basically, the identification of these markers is based on the application of mass spectrometric techniques, which are often proceeded by a chromatographic separation step. This work reviews the most significant results obtained worldwide by the application of analytical procedures based on mass spectrometric techniques to ancient samples such as amphorae, ointment jars and mummies, permitting a critical interpretation and evaluation of the obtained data.

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OL-44 FINAL RESULTS OF THE EUROPEAN RESEARCH PROJECT DIFFERENCE: NEW ALTERNATIVE METHODS FOR DIOXIN ANALYSIS Jacob de Boer, Stefan van Leeuwen, Peter Korytar

Netherlands Institute for Fisheries Research, Wageningen University, P.O. Box 68, 1970 AB IJmuiden, The Netherlands

The European research project DIFFERENCE (“Dioxins in Food and Feed – Reference methods and New Certified Reference Materials”) was focussed on the development of alternative methods for the analysis of polychlorinated dibenzodioxins (PCDDs), dibenzofurans (PCDFs) and dioxin-like polychlorinated biphenyls (dl-PCBs) using comprehensive multi-dimensional gas chromatography (GC×GC), gas chromatography combined with low resolution ion-trap mass spectrometry (GC-LRMS/MS), the CALUX bioassay and an Ah-PCR technique. GC combined with high resolution mass spectrometry (HRMS) was used as a reference method in all comparisons. Given the need for a regular monitoring of PCDD/Fs and PCBs in Europe, which was further enhanced by the implementation of the maximum residue levels (MRLs) for PCDD/Fs in food and animal feed per 1 July 2002, and the relatively high costs of GC-HRMS and the limited capacity for HRMS analyses in European laboratories, cheaper, faster, but reliable methods were badly needed. The method development part included a developmental phase, a validation phase and a standardisation phase. In addition to the method development, attention was also paid to alternative extraction and clean-up methods, in particularly on accelerated solvent extraction. A feasibility study on the preparation and certification of reference materials was also included.

Three out of the four methods have successfully been developed and validated. CALUX may be considered as a valuable screening method that can be used in times of crisis for a large number of samples to indicate if those samples are below, around of above the EU MRLs. Quantitative total-TEQ values may also be produced by this method, but due to the variability of those data, in particular for the mono-ortho substituted PCBs, the use of CALUX in that way is not recommended. The application of recovery correction appeared to be essential in any case. GC-LRMS/MS may be used as a reliable routine method that will produce congener-specific data. GCxGC-ECD or GCxGC-ToF-MS emerged as viable routine methods for dl-PCBs and PCDD/F measurement. No doubt for environmental samples, and for food and feed materials in which dioxins and dl-PCBs occur at higher concentrations (around the MRLs in fatty fish, fish oil), GCxGC is able to serve as an excellent routine method and alternative for HRMS. As soon as dioxin and dl-PCB concentrations drop to low levels, i.e. lower than the MRLs and action levels, a lot of time is needed to produce congener-specific data, to integrate the chromatograms, to adjust the baseline, to compare relative retention times, etc. However, even then GCxGC may still serve as a screening method. GCxGC-ToF-MS is an improvement compared to GCxGC-ECD, but obviously the instrument is considerably more expensive. The fourth method that was studied, an Ah-PCR analysis, is still under development. It is not excluded that that technique may finally also result in a suitable alternative screening method.

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OL-45 MICRO-RAMAN SPECTROSCOPIC INVESTIGATION OF LEMNA MINOR: IMPLICATIONS FOR ENVIRONMENTAL MONITORING APPLICATION

Erzsébet Horváth1, Katalin Kristóf2, Nóra Kováts1

1University of Veszprém, Department of Environmental Engineering and Chemical Technology, H-8200 Veszprém, Egyetem 10. 2Semmelweis University Budapest, Faculty of Medicine, H-1085 Budapest, Üllői 26.

Currently ecological risk assessment is performed by using several standard and non-

standard ecotoxicological tests, which appraise the predicted toxic effect of contaminants. Organisms used in such tests share some common parameters. They represent a critical group (generally a trophic level) in an ecological system exposed to actual contamination. The result of these tests is a quantitative parameter such as mortality or growth inhibition. Lemna minor is a relatively novel test organism recommended by both EPA and OECD. The test parameter is the inhibition of growth, which allows investigation of sublethal toxic effects, as well. Whereas the inhibitory effect of toxins is an empirical parameter, the molecular and physiological bases of this phenomenon are not yet known.

We have investigated the toxicity of Cr(VI) solutions of different concentrations and that of a commercial fungicide, “propisochlor”, on Lemna minor by using micro-Raman spectroscopy. It was found that organic and inorganic contaminants act in different ways. Organic toxins intercalate in the lipid double-layer of the cell membrane, while inorganic ions are transported by ion pumps or channels. Repair mechanisms are active as long as 3-4 days, depending slightly on the concentration of the toxin in question.

Environmental stress effects result in histological changes that can be described quite well by spectral changes. Consequently, micro-Raman spectroscopy minimizes the insecurity of ecological tests based on empirical parameters. Due to the expensive instrumental background, Raman spectroscopy cannot compete with routine ecological tests. However, it helps to understand the physiological response to environmental stress, to find or develop the most relevant (appropriate) test organism, and can also contribute to a better understanding of some properties of biological membranes. [1] Gessner, R., Rösch, P., Kiefer, W., Popp, J.: Raman spectroscopy investigation of biological materials by use of etched and silver coated glass fiber tips. Biopolymers, 67 (4-5), 327-330, 2002. [2] Helmenstine, A., Uziel, M., Vo-Dinh, T.: Measurement of DNA adducts using surface-enhanced Raman spectroscopy. Toxicol Environ Health, 40 (2-3), 195-202, 1993. [3] Wang, Y. N., Galiotis, C., Bader, D. L.: Determination of molecular changes is soft tissues under strain using laser Raman microscopy. Journal of Biomechanics, 33, 483-486, 2000. [4] Pappas, D., Smith, B. W., Winefordner, J. D.: Raman spectroscopy in bioanalysis. Talanta, 51, 131-144, 2000. [5] Naumann, D., Keller, S., Helm, D., Schultz, Ch., Schrader, B.: FT-IR Spectroscopy and FT-Raman Spectroscopy are powerful analytical tools for non-invasive characterization of intact microbial cells. Journal of Molecular Structure, 347, 399-406, 1995. [6] Schrader, B., Klump, H. H., Schenzel, K., Schulz, H.: Non-destructive NIR FT Raman analysis of plants. Journal of Molecular Structure, 509, 201-212, 1999.

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OL-46 LEVELS OF ENVIRONMENTAL POLLUTANTS IN HUMAN BLOOD AND SERUM FROM ITALY Silvano Focardi, Cristiana Guerranti, Guido Perra, Davide Baroni, Ilaria Bisogno, Teresa Benincasa, Aldo Bianchini, Alessandra Schiavone, Nicoletta Ademollo, Francesca Borghini Dipartimento Scienze Ambientali “G. Sarfatti”, Via Mattioli 4, 53100 Siena, Italy Persistent organic pollutants (POPs) and non essential elements are widespread pollutants known to be toxic for biological systems. These substances of low biological degradability tend to biomagnify and bioaccumulate throughout trophic levels. High concentrations have been detected in the tissues of many organisms, particularly predators which include humans, and are known to induce toxic effects. The mechanisms and pathways leading to the presence of such chemicals in human blood are not well characterized but likely involve environmental and dietary exposure. A number of POPs, including PBDEs and dioxins have been identified as endocrine disruptors because they can interfere with hormones causing damage to the vital functions of biological systems. Human exposure to several groups of contaminants was investigated in the general population of Italy. Whole blood and serum samples were collected and analyzed for polychlorinated biphenyls (PCBs), chlorinated pesticides, polychlorinated dibenzo dioxins (PCDDs), polychlorinated dibenzofurans (PCDFs), polybrominated diphenyl ethers (PBDEs), polycyclic aromatic hydrocarbons (PAHs) and heavy metals (Pb, Cd and Hg). Blood has been proven to a suitable matrix for several reasons: it is considered a non destructive method and because it provides a good estimate of the body burden since it is in chemical equilibrium with the organism’s tissues and organs. Contaminant levels in the blood also reflect a wide variety of contamination sources, such as water, air and food, supplying an integrated datum of uptake either via ingestion, inhalation or dermal absorption. All of the above groups of contaminants have been detected in the samples analyzed.

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OL-47 A MONITORING PROGRAM FOR PESTICIDE CONTAMINATION IN SURFACE WATERS IN HUNGARY Erik Maloschik,1 E. Benfenati,2 A. Ernst,1 A. Székács1

1 Plant Protection Institute of the Hungarian Academy of Sciences, Department of Ecotoxicology and Environmental Analysis, H-1525 Budapest, Herman Ottó út 15., Hungary; E-mail: aszek@nki.hu 2 Pharmacological Research Institute “Mario Negri”, Department of Environmental Health Sciences, Via Eritrea 62, 20157 Milan, Italy A five-year survey of pesticide active ingredients and residues in Hungarian surface water samples was carried out within the frame of a national monitoring program. On the basis of physicochemical and ecotoxicological properties of presently registered pesticide active ingredients, the range of analytes was selected to include acetochlor, atrazine, carbofuran, diazinon, fenoxycarb, metribuzin, phorate, prometryn, terbutryn and trifluralin. Target analytes were monitored by GC-MS from 389 surface water samples collected at specific reference points. Samples were prepared for analysis by SPE and SPME. Newly discovered ecotoxicological problems (e.g., endocrine disrupting effects) related to pesticide residues as well as the tightening legal regulations regarding pesticide residues in drinking water (in correlation with Hungary joining the European Union) require analytical methods of increased sensitivity for monitoring pesticide residue levels. In attempt to comply with this requirement, novel bioanalytical methods are being developed and instrumental and immunoanalytical methods are being applied at the Department of Ecotoxicology and Environmental Analysis at Plant Protection Institute of the Hungarian Academy of Sciences to detect and quantitatively monitor pesticide active ingredients and residues. Target analytes in these studies include presently registered and potentially water-contaminating pesticide active ingredients. These substances are being detected partly by gas chromatography – mass spectrometry (GC-MS) and partly by enzyme-labelled immunosorbent assays (ELISAs), while their aqueous toxicity is being measured by the Daphnia magna biotest. In the scope of a five-year monitoring study, sample preparation methods and a GC-MS analytical protocol have been adapted and optimized for such monitoring purpose. This work has been carried out in a close collaboration with the Plant and Soil Conservation Service of the Hungarian Ministry of Agriculture and Regional Development. To provide sample preparation appropriate for GC-MS determinations, solid phase extraction (SPE) and solid phase microextraction (SPME) methods have been applied. GC-MS determinations have been carried out in electron impact (EI) and chemical ionization (CI) modes detecting total ion count (TIC) or in selective ion monitoring (SIM) mode. Detected water contaminants of pesticide origin have been ranked, graphically summarized by geograpical location, correlated with plant protection technologies used in the sampling region. Contamination sources have been estimated, and recommendations to necessary steps to eliminate the contaminants in outstanding point sources of contamination and evaluate the effects of these given contaminants have been formulated. Contamination levels have been ranked into three classes: (1) strongly contaminated sites (> 10,000 ng/l), 2 sampling sites, 2.2% of all sampling locations, clearly point sources of contamination, not of agricultural but industrial origin; (2) contaminated sites (1000 – 10,000 ng/l), 14 sampling sites, 15.6% of all sampling locations, mostly in the Balaton and Northern Hungary regions; (3) slightly contaminated sites (< 1000 ng/l), 32 sampling sites, 35.6% of all sampling locations, even

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distribution with presumably non-point sources of contamination, presenting a continuous environmental stress factor. Thus, determinations indicated pesticide residues over 10 ng/l in 53.4% of all sampling locations, which is a rather large proportion relative to previous expectations. An even more severe factor was that over 90% of the collected surface water samples contained detectable levels of one or more pesticide active ingredients in the following bear-down by active ingredients: acetochlor 30.6%, atrazine 44.4%, diazinone 64.8%, prometryn 17.6%, terbutryn 2.8%, while carbofuran, fenoxycarb, metribuzin, phorate and trifluralin have not been found in any water sample. In another part of the monitoring study, in-house developed ELISA systems have been applied to monitor some of the above pesticide active ingredients. Studies demonstrated that the developed ELISA systems are well applicable for screening of water samples for pesticide residues prior to GC-MS determinations.

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OL-48 ANALYSIS OF SOME ELEMENTS IN FOUR TYPICAL FOODS FROM WUHAN

Jun Yao1,*, Atakora Djah1, Jian-chan Wu1, Hong-yin Han1, Gyula Záray2

1 School of Environmental Studies, China University of Geosciences, Wuhan, Hubei province, 430074, P. R. China 2 Department of Inorganic and Analytical Chemsitry, Eötvös University, Budapest, P.O. Box 32. H-1518, Hungary

This paper as a part of work in Sino-Hungarian international science and technology cooperation project, we have found a analytical method to determine the inorganic and organic form of eighteen heavy metals in typical foods from Wuhan by separate technology and atom spectrometry analysis. Concentrations of eighteen elements (As, Ba, Ca, Cd, Co, Cr, Cu, Fe, K, Li, Mn, Na, Ni, Pb, Sb, Se, Sr, Zn) were determined by HPLC-ICP-AES and LC-ICP-MS. To evaluate the elements content in seasonal food, we choose same food but in different seasons. According to the data got from the experiment, we summed up and analyzed them in three different ways: a) Compare the elements concentration of four different species in the same season(summer); b) Compare the elements concentration of seasonal cabbage; c) Compare the maximum concentration of elements in different species. The results showed that concentrations of same elements are obviously different in different seasons. For the essential elements Ca, Cu, Mg and for the nonessential and toxic elements Ba, As, Cd, Pb and Sb, this difference was highly significant. In spring, some elements in cabbage had much lower concentration than in other seasons. For example, As:0.31µg/g; Ni:2.14µg/g. On the contrary, some elements had much higher concentration, for example, Ba:13.5µg/g; Cr:1.05µg/g; Cu:6.73µg/g; Li:2.15µg/g; Na:1993µg/g; Pb:5.38µg/g. Moreover, the concentration of some elements were higher in cabbage than that in others (As:1.62µg/g; Ba:13.5µg/g; Cd:0.92µg/g; Co:0.34µg/g; Mn:55.5µg/g; Sr:112.3µg/g) On the contrary, the content of other elements for cabbage was lower (Cu:1.7µg/g; Na:1993µg/g).

We also set up advanced evaluation system to evaluate the impact made on people’ health by some toxic metals, so as to evaluate the relationship among heavy metals’ exposure, organisms and people’ accident.

Keywords: HPLC-ICP-AES; LC-ICP-MS, essential element, toxic element; food Acknowledgements: We gratefully acknowledge the financial support of Sino-Hungarian International Science and Technology Cooperation Project; National Natural Science Foundation of China (No. 30170010); and Scientific Research Foundation for the Returned Overseas Chinese Scholars, China University of Geosciences, Wuhan (XLX04901).

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THURSDAY, 27 OCTOBER 2005

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OL-49 RoHS DIRECTIVE – HOW TO DO THE RELATED ANALYSIS László Bezur Budapest University of Technology and Economics, Institute for Gereral and Analytical Chemistry, 1111. Budapest, Szt. Gelléret tér 4; E-mail: bezur@mail.bme.hu

Directive 2002/95/EC of the European Parliament of the Council of 27 January 2003 on the restriction of the use of certain hasardous substancens in electrical and electronic equipment, commonly known as RoHS (from Restrictiom of Hasardous Substances, becams effective July 1, 2006. The RoHS directive mandates legislation restricting the chemical content levels of certain hazardous compounds that may occour in electrical and electronic equipment. The requirement’s purpose is the protection of health by promoting environmentally sound recovery and disposal of Waste electrical and electronic equipment within the European Community. Specifically, it indentifies and reduces the content of certain metals and hasardous compounds based on scientific evidence and studies. The Directive minimizes potential differences between the laws of Member states. The directive acknolwledges that the identified compounds will be difficult to eliminate from certain applictions and substitution materials must be developed. Therefore, certain exemptions are listed in an annex. These are to be reviwed every four years and additional hazardous materials may also be added. Some of the current exemption include mercury in various fluorescent and certain other lamps, lead in glass or alloys of steel, lead in solders in network infrastructure equipment and chromium(VI), for anti-corrosion of steel, in cooling systems of absorption refrigerators. The current hazardous compounds and their limit by weight are as follows: (i) lead 0.1%, (ii) mercury, 0.1%, cadmium, 0.01%, chromium(VI), 0,1%, Polybrominated Biphenyls (PBBs), 0,1%, Polybrominated Diphenyl Ethers (PBDEs), 0,1%. Producers of equipment are held responsible for ensuring that their products do not contain the six restricted substances. Equipment producers will need to abtain materials declarations or certificates of complience from their suppliers or they have to carry out the analysis of the component for the restricted components. In our present work a such analytical study was carried out. The complience of a CD drive unit was tested so that all the components and sub units have been analysed individualy. The test was limitet to the restricted metallic components, Pb, Hg, Cd, Cr(VI). Different sample preparation methods have been developed followed by ICP-OES, Flame-AAS and CV-Hg-AAS analysis of the sample solutions.

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OL-50 THE ACTIVITIES OF THE WORKING GROUP ON GOOD LABORATORY PRACTICE OF THE ORGANISATION FOR ECONOMIC CO-OPERATION AND DEVELOPMENT

Sergio Caroli

Istituto Superiore di Sanità, Rome, Italy; E-mail, caroli@iss.it. In the late 1970’s control of chemical substances in the international trade raised much concern in most industrialised countries because of the adverse effects potentially toxic chemicals could have on human health and the environment. To properly deal with this issue, the Organisation for Economic Co-operation and Development (OECD) launched a number of programmes aimed at providing decision makers with reliable experimental information on new chemical substances so as to allow for a sound assessment of the benefit-to-risk ratio well before chemicals are produced and marketed. The cornerstone of this policy is the implementation of the so-called principles of Good Laboratory Practice (GLP) whenever non-clinical safety studies on new chemicals are undertaken by Test Facilities (TFs). The OECD Working Group on GLP, established in the early 1980’s, is a permanent forum with representatives of about 40 countries interested in the mutual recognition of studies performed according to the GLP principles because of the ensuing obvious beneficial consequences, i.e., saving of financial resources and better protection of human health and the environment. An overview is presented of the major activities currently carried out by the OECD Working Group on GLP in this context, namely: i) updating of the GLP principles to take into account new application fields (e.g., in vitro tests and archiving procedures); ii) conduct of Mutual Joint Visits to verify the way GLP Monitoring Authorities discharge their duties in their own country; iii) organisation of Training Courses for GLP inspectors in order to harmonise as much as possible the various approaches to GLP inspections; iv) interaction with the ISO world to explore the possibility of joint activities in fields common to the GLP and ISO/IEC 17025 quality systems (e.g., analytical tests and validation of analytical methods).

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OL-51 APPLICATION OF EU WATER FRAMEWORK DIRECTIVES IN MONITORING OF SMALL WATER CATCHMENT AREAS IN HUNGARY I. SELECTION AND CHARACTERIZATION OF A REFERENCE SITE FOR SMALL CREEKS IN MÁTRA MOUNTAINS

Balázs László1, Andrea Zagyva2, György Heltai3 1Budapest University of Technology and Economics, Department of Sanitary and Environmental Engineering 2Environmental Protection and Water Management Research Institute, Institute for Water Pollution Control and Nature Conservation 3Szent István University, Department of Chemistry and Biochemistry The EU Water Framework Directives (WFD) prescribe to achieve a good ecological quality of surface water systems for Hungary by 2015. To keep this deadline an establishment of a new three level (surveillance, operational and investigative) ecological monitoring system is required, including hydrological, biological and chemical components. The EU WFD does not prescribe any standard for the human water uses. The ecological status should be related to the reference conditions of a water body, having a certain type. Up to now in Hungary the standard network of surface water quality monitoring has been operated for rivers and large lakes, but no satisfactory information was available about small water bodies. Therefore, there is an urgent need to design a methodology for developing monitoring network of such surface waters systems. For this purpose a slightly impacted mountain creek (Nagy Creek in Mátra mountains was selected and closely monitored during one year (May 2004 →May 2005). In this program hydrological, biological and chemical parameters required by WFD were monitored. Variability of parameters vs. time and site was also carefully investigated. The minimum frequency of sampling was determined in time and space for selected ecological components (physico-chemical compounds and phytoplankton) to meet the WFD standards for confidence. It was found that the minimum frequency of sampling, suggested by the WFD is far less than that of the necessary one.

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OL-52 APPLICATION OF EU WATER FRAMEWORK DIRECTIVES IN MONITORING OF SMALL WATER CATCHMENT AREAS IN HUNGARY II. CHARACTERIZATION OF THE MODERATELY LOADED, RURAL CATCHMENT AREA OF GALGA STREAM Emőke Szlepák1, György Heltai1, Eszter Szilágyi2, Béla Csányi3, Emőke Sz. Bardóczy4

1 Szent István University, Department of Chemistry and Biochemistry 2 Budapest University of Technology and Economics, Department of Sanitary and Environmental Engineering 3 Water Resources Reserach Plc., Institute of Water Pollution Control 4 Szent István University, Deparment of Landscape Ecology

The EU Water Framework Directives (WFD) prescribe to achive a good ecological quality of surface water systems for Hungary by 2015. To keep this deadline an establishment of a new three level (surveillance, operational and investigative) ecological monitoring system is required, including hydrological, biological and chemical components. The EU WFD does not prescribe any standard for the human water uses. The ecological status should be related to the reference conditions of a water body having a certain type. Up to now in Hungary the standard network of surface water quality monitoring has been operated for rivers and large lakes, but no satisfactory information was available about small water bodies. Therefore there is an urgent need to design a methodology for developing monitoring network of such surface waters systems. For elaboration of a monitoring guidance, selection of model catchment areas with different level of anthropogenic impacts was necessary. For characterization of a moderately loaded, rural type catchment area, the mid-altitude and lowland type river basin area of Galga stream was selected. Hydrological, biological and chemical parameters required by WFD were closely monitored in this project during one year (May 2004 →May 2005). Variability of parameters vs. time and space was also carefully investigated. The required minimum frequency of sampling was determined for selected ecological components (physico-chemical compounds and phytoplankton) to meet the WFD standards for confidence. On the basis of the results it should be concluded that the diffuse pollution sources are characteristic for such type of area, however, point sources can be identified, too. This character proved an opportunity for studying the movement of a point source heavy metal pollution with the sediment.

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OL-53 APPLICATION OF EU WATER FRAMEWORK DIRECTIVES FOR MONITORING OF SMALL WATER CATCHMENT AREAS IN HUNGARY III. STREAMS IN HEAVILY LOADED URBAN AREA Gábor Halász1, Ilona Fekete1, Eszter Szilágyi2, Béla Csányi3, György Heltai1 1Szent István University, Department of Chemistry and Biochemistry 2Budapest University of Technology and Economics, Department of Sanitary and Environmental Engineering 3Water Resources Research Plc., Institute of Water Pollution Control

The EU Water Framework Directive (WFD) prescribe to achieve a good ecological quality of surface water systems for Hungary by 2015. To keep this deadline an establishment of a new three level (surveillance, operational and investigative) ecological monitoring system is required, including hydrological, biological and chemical elements. The WFD does not prescribe any standards for human water uses. The ecological status should be related to the reference conditions of a water body, having a certain type. Up to now, water quality monitoring network of surface waters has been operated for rivers and larger lakes in Hungary, but no satisfactory information was available for small waters. Therefore, there is an urgent need to develop an appropriate methodology for monitoring network of these water bodies. For elaboration of a monitoring guidance, catchment areas with different level of anthropogenic impacts were selected. Among these, the most heavily loaded area is the Rákos stream with its artificial lake system between the towns of Gödöllő and Isaszeg. The Rákos stream originates in Gödöllő and reaches the Danube in Budapest through an artifically covered bed exposed to many urban and industrial impacts. Hydrological, biological and chemical parameters required by WFD were monitored in this project during one year (May 2004 →May 2005). Variability of parameters vs. time and space was also carefully investigated. The required minimum frequency of sampling was determined for selected ecological components (physico-chemical compounds and phytoplankton) to meet the WFD standards for confidence. It was found that the minimum frequency of sampling, suggested by the WFD is far less than that of the necessary one. On the basis of the above monitoring program the necessary actions were determined to meet the good ecologycal status or potential of the water bodies. Investigative and operational monitoring network was also designed.

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OL-54 INFLUENCE OF C/N RATIO ON NOX AND CO2 PRODUCTION OF CULTIVATED SOIL IN CLOSED MICROCOSM SYSTEM

Krisztina Kristóf1, Abdousalam A. Algaidi1, Györgyi Kampfl1, Hosam Bayoumi2, György Heltai1

1Department of Chemistry and Biochemistry, Szent István University, H-2103 Gödöllő 2Ph.D. Environmental Sciences Institute, Szent István University, H-2103 Gödöllő

It is important to study the impact of denitrification and nitrification in soil ecosystems

on the emission of gases (N2O, CO2 and NO) whose increasing amount in the atmosphere may cause serious environmental problems e.g. global climate change and ozone depleting. In the year of 2002 and 2003 the CO2 and N2O production from cultivated soil and the effect of different factors on the gas emission were studied. In 2004 and 2005 further two microcosm experiments were carried out to confirm and to complete the previous results. The experiments were conducted using closed glass vessels, brown forest soil, KNO3 fertiliser, maize residue and water in the same way as it was applied in 2003. The C/N ratio was set by KNO3 and the organic additive. During the measurement, the vessels were stored in a thermostat at 15 °C in 2004 and at 28 °C in 2005. The N2O and CO2 content of the head space gas samples were analysed by a gas chromatograph and the NO concentration was measured by a chemiluminescence NO-analyser in both experiments. In the beginning and in the end germ count was also performed to demonstrate the presence of microbial activity.

From the results of gas content measurements and germ count it could be concluded that while the organic matter supply was increasing microbial activity resulting in higher CO2 concentration, the use of N-fertiliser was decreasing it. These results were consistent with the results of the previous experiments conducted e.g. in 2002 in case of N2O and CO2. In 2005 due to the higher temperature the tendencies which were also detected earlier became more definite. It became clear that the gas concentrations changed in time according to maximum curves. Although the amount of CO2 and N2O was higher than previously, in accordance with the former results the rate of the N2O-N loss was still lower than it is published from similar experiments. Surprisingly the amount of NO production was not influenced by the higher temperature. The change of NO concentration in time showed a maximum curve as well which was explained by the short lifetime of the gas. The organic matter addition delayed the appearance of maximal gas concentration. To clarify the underlying processes and to confirm the present results further studies are required.

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ABSTRACTS OF POSTER PRESENTATIONS

TUESDAY, 26 OCTOBER 2005

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P01 COLOUR FADING IN TEXTILES: AN INSIGHT ON DEGRADATION PRODUCTS A. Andreotti, M. P. Colombini, I. Degano, J. Lucejko

Department of Chemistry and Industrial Chemistry, University of Pisa, Via Risorgimento, 35, 56126 Pisa, Italy

Studying the materials used in textile dyeing may help considerably in understanding

how the object itself originally looked, and in dating and locating its origin. It may also be valuable in deciding how it should be conserved. Since ancient times many natural materials were used in Europe. The most common yellow dyes were extracted from weld (Reseda Luteola L.), young fustic (Cotinus Coggyria Scop.), dyer’s greenweld (Genista Tinctoria L.), sawwort (Serratula Tinctoria L. Gaud.), and the berries of some species of Rhamnus. Later on, old fustic (Chlorophora Tinctoria L. Gaud.) was introduced from the West Indies, and in the 18th century quercitron bark (Quercus velutina L.) began to be imported from North America. Soluble redwood dyes, extracted from various species of the genus Caesalpina, were also used.

Identifying these dyes in old textiles is difficult due to the poor lightfastness of their flavonoid constituents and the complexity of the degradation processes, which is not yet completely understood.

This paper deals with the application of GC-MS and DE-MS methods to identify flavonoids in raw plant materials (weld, dyer’s greenweld, Buckthorn berries, onion skins, old fustic) and in fresh and aged alum-mordanted wool dyed specimens. The ageing treatment consists of exposure to artificial daylight (lamp with outdoor filter at 280 nm) at room temperature and 50% RH.

The method allows the easy identification of a dyestuff by the detection of the molecular markers surviving in old textiles and reproduced in fresh materials under accelerated ageing.

In particular, a procedure for the extraction and derivatisation of flavonoids with N,O-Bis(trimethylsilyl)trifluoroacetamide in order to make them detectable in complex mixtures by GC-MS analysis has been developed. The DE-MS technique was used to detect compounds at a 1µg/g level with a minimum sample treatment in plant and wool extracts. The technique was also employed to characterize the fractions collected from an HPLC.

The most significant results on fresh and aged samples are discussed.

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P02 ARCHAEOLOGICAL WOOD CHARACTERISATION BY NMR AND PY/GC/MS

M. Orlandi1, M. P. Colombini2, F. Modugno2 M. Sardelli2, E-L. Tolppa1, L. Zoia1

1Università di Milano-Bicocca, Milano, Italy 2Dipartimento di Chimica e Chimica Industriale, Università di Pisa, Italy

At present, few information is available on the chemical transformation undergone by wood in the course of degradation processes occurring in archaeological and artistic objects. This lack of information also covers the specific case of wet wood. Considered that in degraded wood, especially after long permanence in humid/wet environment, the cellulose content is generally very low in respect to lignin, the characterisation of lignin is an aspect of primary interest.

In this paper, archaeological waterlogged wood samples and lignin samples extracted from archaeological waterlogged wood have been characterised by mean of pyrolysis coupled with mass spectrometry (Py/GC/MS), in order to investigate the potentialities of these analytical techniques in the characterisation of archaeological wood. Organic extractives of the same samples have been also characterised by gas chromatography/mass spectrometry (GC/MS). The results have been compared with those obtained for unaged wood and lignin. The characterization of chemical structure of acetylated lignin extracted from archaeological waterlogged was assessed by heteronuclear single quantum coherence (HSQC) and 13C- NMR spectroscopy.

The chemical study of the modification involving lignin in the course of degradation can contribute not only to a better knowledge of the degradation processes of wood in artistic and archaeological objects but also to the development of consolidation and conservation procedures.

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P03 CHARACTERISATION OF VEGETABLE OILS IN ARCHAEOLOGICAL FINDINGS Maria Perla Colombini, Francesca Modugno, Erika Ribechini Department of Chemistry and Industrial Chemistry, University of Pisa, Via Risorgimento, 35, 56126 Pisa, Italy

Archaeological sites often bring to light a wide variety of objects and materials which have been collected, processed and used by humans over the centuries. Due to their long-standing burial, some of these materials and objects, especially those of organic nature, have been partially or totally altered. Although the preservation of organic material is still considered quite unusual in archaeological environments, at the end of the 1980s, analytical investigations began to provide evidence of the widespread survival of organic residues, and highlighted a very high level of preservation in several cases. Consequently, the identification of specific materials from the molecular patterns and from peculiar biomarkers can help in assessing the role that these substances played, and in determining the use of artefacts.

A wide range of natural substances may be found in association with archaeological remains. Of these, vegetable oils are known to have been used extensively by ancient populations in cookery, for lighting, and as ingredients for cosmetics, balms and medications.

This paper presents a GC/MS study on the chemical transformations of some vegetables oils (olive oil from Olea europaea, palm oil from Elaeis guinnesis, coconut oil from Cocos nucifera, safflower oil from Carthamus tinctorius, castor oil from Ricinus communis, almond oil from Amygdalus communis, brassica oil from Brassica juncea, and moringa oil from Moringa oleifera), when subjected to induced degradation processes by means of thermo-oxidative treatments.

The most significant results on the identification of the molecular markers detected after the accelerated ageing are given, together with their utility for assessing the origin of a vegetable oil in archaeological residues. Finally, the results of the analysis of organic residues present in archaeological findings from Pompeii (Italy) are discussed.

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P04 EFFECT OF ALKALI METAL CONTENT ON WOOD ASH IN FLUID BED BOILER. INVESTIGATION OF ADHESION PROPERTY OF BED MATERIAL OBTAINED IN WOOD BURNING FLUID BOILER Éva Bandi1, Lívia Nagy2, Linda Sárga3, Géza Nagy2,3

1PannonPower Holding Ltd Pécs, H-7632 Pécs, Edison út 1; E-mail: kalorikus@pannonpower.hu 2Hungarian Academy of Sciences, Research Group for Chemical Sensors, H-7624 Pécs, Ifjúság útja 6 3University of Pécs, Department of General and Physical Chemistry, H-7624 Pécs, Ifjúság útja 6

The global need for reduction of carbon dioxide emission is well known. The fossil fuel based energy production is one of the major sources of atmospheric CO2. One way of decreasing CO2 emission is substitution of fossil fuel with wood and other biomass. In order to achieve efficient utilisation of biomass for energy production, several technological problems have to be solved. Sometimes, very small differences in fuel composition result in different requirements concerning the structure of the boiler, the temperature operation, etc. The low temperature melting ash can form glassy depositions on the heat exchanger surfaces causing loss of efficiency and corrosion.

In our study, a comparison between wood ash composition, burning conditions and thermal behaviour has been carried out. In the experiments, TGA, DTA thermoanalytical measurements were carried out in the range of 30 – 850 °C. Moreover, the K, Na, Ca and Si contents were determined by atomic spectroscopic analysis (FES, AAS). The burning conditions were characterised. Wood ash samples were taken from the bed material and from the deposited glassy melts.

In the poster, the results obtained will be summarised together with the sample preparation and other details of the experiment. The optimal burning condition for the different biomasses studied could be determined, which provided effective energy production and “problem free” combustion without the formation of glassy deposition.

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P05 STUDY OF ATOMIZATION AND GENERATING PROCESSES WITH THE FES/AAS COMBUSTION SYSTEM ENSURING 100% SAMPLE INPUT Áron Béni1, Ildikó Kiss 1, Sándor Berényi 2, József Posta2 1Department of Inorganic and Analytical Chemistry, University of Debrecen, Egyetem tér 1., H- 4032 Debrecen, Hungary. E-mail: postaj@tigris.klte.hu 2Department of Agricultural Chemistry, University of Debrecen, Böszörményi út 138., H-4032 Debrecen, Hungary One of the fundamental factors of the productive capacity of atom spectroscopic methods is the efficiency of sample intake into the flame and plasma. By utilisation of the formerly developed hydraulic high-pressure nebulizer [1-2], we have worked out an nebulizer/burner system with a sample intake efficiency of 100%. The system is constructed such that the hydraulic high-pressure nebulizer (HHPN) is positioned coaxially in the interior of the Meker-burner manufactured for the atom absorption spectrometer. The nebulizer is positioned in the centre of the burner with suitable distance pieces, so that the high-pressure liquid jet from the nebulizer passes unobstructed through the central aperture of the 13 circular apertures of 2 mm in diameter. The high-pressure liquid jet emerging from the burner nebulizer, impacting with the ceramic rod positioned in the flame is vaporized directly in the high-temperature area where atomisation of the sample takes place. The flexible PEEK tube, connecting the high-pressure pump and the nebulizer is passed through the plastic plug in the rear wall of the spray-chamber with airtight sealing. We have compared the particle distributions obtained with the new sample intake method and the traditional pneumatic sample intake. Optimizing operation parameters of the nebulizer/burner system, an improvement of 2-12 time was achieved for the signal/noise relation in the flame emission and flame atom absorption spectrometry with sample input under the new system dependent on nebulization rate. Similar results were achieved in the case of chromium(III) and chromium(VI) on-line chromatographic separation [3]. This nebulizer is extremely suitable for development of liquid chromatography – AAS/FES coupled technology for specialized analytical tasks. The nebulizer/burner system can easily be put into operation with small conversions to the commercial atom absorption spectrometers. Besides its favourable analytical features, it can be used extremely well for the development of coupled technology in specialized analytics, since it ensured the best analytical sensitivity with the small liquid flows applied in efficient HPLC chromatographic separation. [1] H. Berndt, Fresenius Z. Anal. Chem., 331, 321 (1988) [2] J. Posta, H. Berndt, Spectrochim. Acta, 47B, 993 (1992) [3] A. Gáspár, Cs. Sógor, J. Posta, Fresenius J. Anal. Chem., 363, 480 (1999)

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P06 FOLLOWING OF “IN VITRO” EXTRACTION OF METALS FROM TABLETS BY ATOMIC ABSORPTION SPECTROMETRY Ildikó Kiss, Lívia Varga, József Posta Department of Inorganic and Analytical Chemistry, University of Debrecen, Egyetem tér 1, H-4032 Debrecen, Hungary; E-mail: postaj@tigris.unideb.hu

Mineral and trace materials are very important in conservation of biological balance of human body. Recently in consequence of the insufficiency of elements in food there is no quantity enough macro- and microelements for preservation of our health, hence different drugs are used to ingest into the human body. The task of our research work was to examine the content of effective substance and their extraction in ten different drugs among ”in vitro” conditions. The rate of extraction and the quantity of extracted metal ions were determined in synthetic gastric acid. The examined drugs are the followings: Abtei, Actival Max, Centrum, Every One, Flu-Vit, Humetta, Mega Minerals, Supherb, Supradyn and Universal Daily One.

The digestion was carried out in a MILESTON 1200 Mega High Performance Microwave Digestion Unit. A Unicam SP 1900 AAS instrument was applied for analysis of elements.

The tablets were taken into synthetic gastric acid to examine the extraction. The solution was sampled in every half an hours with continuous mixing on 37 °C temperature.

The zinc, calcium, potassium, magnesium, manganese, copper and iron were analyzed in the drugs. Generally, there was no significance difference between the declared and analyzed metal content. However, e.g. in the case of Mega Minerals tablet there was no detectable copper content in our analysis.

The extraction of one part of metal ions in synthetic gastric acid is shown similar pattern. The rate of extraction is significant in the first half an hour and moderate in the next an hour and a half, while it increases in the 3rd hour. “This extraction pattern” is characteristic for zinc, iron, potassium, magnesium, manganese and calcium. The extraction of copper is different from the above elements. The rate of extraction in the first half an hour is significant in the case of Actival Max and the quantity of copper is approximately constant in the next two hours and a half. The extraction of copper is not significant in the case of Centrum tablet.

As our examinations were carried out in synthetic gastric acid, thus the possible extraction in the stomach was modelled, however certain drugs did not collapse in synthetic gastric acid (e.g. Mega Minerals), so these drugs will be examined in synthetic intestinal juice.

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P07 DETERMINATION OF CHROMIUM CONTENT IN FOODSTUFFS AND NUTRITION SUPPLEMENTS Róbert Kovács, Áron Béni, Roland Karosi, Csilla Sógor, József Posta Department of Inorganic and Analytical Chemistry, University of Debrecen, Egyetem tér 1., H- 4032 Debrecen, Hungary. E-mail: postaj@tigris.klte.hu

In the course of our work we established the chromium content of foodstuffs, beverages and nutrition supplements. We determined as to in what quantity do these cover the organism’s daily chromium requirement. The obtained results were compared with data published by other authors and we experienced that these data not only differ significantly from the results obtained by us, but from each other as well. We established the total chromium content in cereal milling products, in breads, in certain plants (broccoli, thyme), in eggs and in alcoholic beverages (wines, beers) originating from various sources. Comparing our measurement results with literary data originating from various sources, it turned out that the published data display significant deviations and do not represent the actual chromium content in regard to certain domestic products. We concluded from our examinations that the methods and circumstances of producing the foodstuffs, the utensils and equipment utilised for their production and storage (with particular regard to stainless steel utensils and cans), further the origin of the foodstuffs (according to geographical area of production) influence the chromium content to a large degree. Therefore it is not expedient to gather information from sources where, for example the precise type (e.g. cheese, wines, etc.) and precise mass (e.g. half-one cup of broccoli) of the given foodstuff cannot be found. It can be said that smaller measures of deviation are also displayed in relation to foodstuffs of identical type (e.g. egg-yolk) but originating from different sources, therefore when planning diets serving for supplementing chromium intake, it is questionable as to which source is to be regarded as definitive. Therefore a simpler solution for sportsmen and women could be the consumption of certain diet supplement preparations containing chromium. At present there is no literary data available to us demonstrating whether toxic chromium compounds can occur from the natural Cr(III) content of cereal milling products used when baking and toasting bread, that is why we considered the examination of this to be important also. The thermal experiments showed that hexavalent chromium compounds do not occur when baking and toasting bread, moreover if Cr(VI) should be present in the flour or in the bread prepared from it, then it would reduce to Cr(III). It is revealed from comparing literary rows of data originating from various sources, that if we want to plan our diet in regard to intake of chromium, on the basis of the Internet, then we can make mistakes to an order of magnitude. In every case the chromium content of a given foodstuff can only be decided on the basis of concrete samples and concrete analysis, and unfortunately the obstructs the possibility of people attempting to compile diets themselves, with the help of which they could provide their organisms in known and required quantities. The use of diet supplement preparations containing chromium could offer assistance in solving this problem.

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P08 DEVELOPMENT OF SdFFF COUPLED ON-LINE TO CID-ETAAS FOR THE CHARACTERIZATION OF ENVIRONMENTAL COLLOIDS G. Blo1, A. Ceccarini2, C. Conato1, C. Contado1, F. Dondi1, F. Fagioli1, R. Fuoco2, A. Pagnoni1, A. Passerini1 1Dipartimento di Chimica, Università di Ferrara, via L. Borsari 46, 44100 Ferrara. 2Dipartimento di Chimica e Chimica Industriale, Università di Pisa, via Risorgimento 35, 56126 Pisa

Environmental colloids play an important role both in performing pollutant transport (e.g. in aquatic systems or in the atmosphere) and in enhancing their accumulation (i.e. in soils), depending upon the degree of mobility of the colloids. Many methods have been developed in order to characterize colloidal matter, but the sizing of material in the colloidal range and their chemical characterization is difficult to obtain by a single method. The analytical approach requires a preliminary separation step, in order to obtain relatively monodisperse fractions to present to the detection system for chemical characterization. Field-Flow Fractionation (FFF) methods allow different size fractions (across the sample size distribution) to be collected for subsequent investigation, for example, heavy metal load determination. In particular, the Sedimentation Field-Flow Fractionation (SdFFF) technique, which has already been applied to environmental colloids, is an established method for the high resolution, mass based separation and sizing of colloids in the 0.05 - 1.0 mm size range. A limiting factor of this technique is the small amount of the sample that can be processed in a single FFF separation run. This means that only a very sensitive analytical method can be used to characterize the separated particle fraction; otherwise time-consuming concentration steps are required which entail further sample handling.

In the present work Electro-thermal Atomic Absorption Spectrometry (ET-AAS) is applied as an on-line detection system for SdFFF. The coupling is performed through a pre-concentration device - Capillary Injection Device (CID) - which allows the direct introduction of the analytical solution into the graphite furnace to an extent of some ten millilitres, thus providing an in-situ preconcentration. The hyphenation of these techniques permits the determination of trace elements at pg/g levels, on different size range colloidal fractions, in the SdFFF eluate. It avoids the concentration steps and possible sample contamination. The coupled SdFFF-CID-ETAAS system performance has been investigated by applying the procedure on silica and iron (III) oxide standard particles of a known size distribution.

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P09 ADVANCES IN CLINICAL APPLICATION OF BREATH ANALYSIS BY GAS CHROMATOGRAPHY/MASS SPECTROMETRY F. Di Francesco1, M. Onor2, S. Tabucchi3, M. G. Trivella1, A. Ceccarini3, T. Schäfer3, R. Fuoco3

1Institute of Clinical Physiology, CNR, Via Moruzzi 1, 56100 Pisa, Italy 2Institute for the Chemical-Physical Processes, CNR, Via Moruzzi 1, 56100 Pisa, Italy 3Department of Chemistry and Industrial Chemistry, University of Pisa, Via Risorgimento 35, 56126 Pisa, Italy

Since Hippocrates’ time, physicians have known that human breath might provide sound information on health conditions. If a pathology causes an alteration in the normal metabolic pathways thus giving rise to new compounds, it is reasonable to suppose that these compounds are transported by the blood and released into the alveolar air according to their partition coefficient. At the same time, any compound present in ambient air is, to some extent, dissolved into the blood until an equilibrium is reached; the same compound is then released in breath as soon as its concentration in the air falls. This phenomenon can be exploited to diagnose a pathology or to monitor its course, as well as to assess occupational exposure to toxic volatiles in a non-invasive way. Over 4000 chemicals have been identified in human breath, some of which are related to normal or impaired metabolic processes. Much of the literature in the last century on breath analysis focused on ethanol levels in blood and other forensic topics, or on occupational exposure to solvents. There is only a limited number of studies regarding the most interesting clinical applications and their results are not easy to generalize due to the lack of accepted standard analytical procedures. The low concentration levels of compounds in breath, often in the nmole/l –pmole/l range, together with the presence of bulk interferents make sampling, sample storage and instrumental analysis all very critical. Moreover, insufficient knowledge of many metabolic pathways, both normal and impaired, along with the high intra- and inter-individual variability make it difficult to recognize marker compounds. These reasons may help to explain why breath analysis has failed to enter into common clinical practice.

In this work contributions to both the development of analytical procedures and marker identification are presented. A system for selective water removal from breath samples was set up consisting of a 5 l sampling bag (polyethyleneterephtalate), a PTFE membrane pump, a stainless steel membrane extraction module, and an adsorption tube. Two circuits were connected to the module, in which two spiral channels were separated by the membrane. In the first circuit, the breath sample was recirculated by the membrane pump from the bag into the module and back into the bag, while in the second circuit a helium flow was used to carry the compounds permeating across the membrane into the tube. In this way, moisture in the breath was substantially removed, and this allowed the tube to be cooled without the formation of water condensate and hence the recovery of an increased amount of compounds. Two membranes (a homogeneous polydimethylsiloxane and a composite polyoctylmethylsiloxane membrane) and two sorbent phases (Tenax GR and a multilayer composed of two graphited carbons and a molecular sieve) were tested. On the application side, diabetes and rejection after liver transplant were considered. Diabetes is a widespread metabolic syndrome that may be almost a-symptomatic for a long time, during which a lot of systemic damage is produced. Liver transplant is the main therapy for many hepatoses, and acute and chronic rejection is the main cause of failure. In this work, the most significant

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findings of a preliminary study carried out with patients at different stages of the illness and healthy volunteers as a control group are reported. Terpenes, linear and branched hydrocarbons, carbonyl compounds, alcohols, sulphur compounds, aromatic and other exogenous compounds were determined and preliminary results on the correlation with the pathologies are discussed.

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P10 SCREENING OF ENDOGENOUS CYTOKININS AND INDOLE-3-ACETIC ACID IN NICOTIANA GLAUCA PLANTS BY CHEMICAL DERIVATIZATION AND GAS CHROMATOGRAPHY-MASS SPECTROMETRY ANALYSIS R. Fuoco1, S. Giannarelli1, B. Muscatello1, P. Bogani2, M. Buiatti2, A. Rosati2 1Department of Chemistry and Industrial Chemistry, University of Pisa, Via Risorgimento 35, I–56126 Pisa, Italy 2Department of Animal Biology and Genetics, University of Florence, Via Romana 17, I-50125 Firenze, Italy

IAA (the major native auxin in many plants) and cytokinins are plant hormones which play key roles in growth and development. Analysing phytohormones is particularly difficult because of their very low absolute and relative concentrations in plant extracts. This means that many extraction and purification steps are required prior to quantitative measurement by GC-MS. A method, which is both fast and consistent, is thus needed in order to identify and measure the cytokinins/auxins ratio. This information, along with the dynamic analysis of plant development, should lead to a better understanding of the action mechanism of phytoregulators, whose concentrations may become useful markers of the response to both biotic and abiotic stress conditions.

The method described here consists in selective extraction and purification from plant materials in a single step using mixed-mode cation exchange and reversed-phase column. The phytohormones thereby collected are pure enough to be quantitatively analysed by GC-MS after derivatisation with N,O-bis(trimethylsilyl)trifluoroacetamide (BSTFA). A detection limit as low as 1 picomole injected was obtained with a typical relative standard deviation of 25%.

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P11 IMAGING ELEMENTAL DISTRIBUTION IN THIN CRYOSECTIONS OF SOFT TISSUES BY SIMS I. Marchetti1, L. Menichetti2, G. Tamborini4, C. Kusmic2, C. Cappelletti2, L. Gaetano2, G. Lazzerini2, R. Fuoco1, P. A. Salvadori2, A. L’Abbate2,3, M. Betti4

1Department of Chemistry and Industrial Chemistry, University of Pisa, Via Risorgimento 35, I-56126 Pisa, Italy 2C.N.R. Istitute of Clinical Physiology, Via Moruzzi 1, I-56100 Pisa 3Sant'Anna School of Advanced Studies, P.za Martiri della Libertà 33, I-56127 Pisa, Italy 4European Commission, Joint Research Centre, Institute for Transuranium Elements, Karlsruhe, P.O. Box 2340, D-76125 Karlsruhe, Germany Secondary ion mass spectrometry (SIMS) is the only mass spectrometry technique able to image the isotopic composition of cells and tissues with spatial resolution of ~1µm. This ion microscopy modality is based on the bombardment of an ion beam a sample and the subsequent production of secondary ions from the sample surface with typical pattern representing the surface chemistry of the specimen. SIMS is highly sensitive (ppm – ppb) and can provide multi-elemental images and quantification: the only important requirement is the electrical conductivity of the sample/sample holder. Biological samples need to be placed on a conductive substrate like indium or silicon and, as far as sample preparation is concerned, cryo-techniques are needed in order to maintain the native ion distribution. Nowadays isotopic imaging with subcellular informations, broad elemental sensitivity, multi-isotopic (elemental) imaging from the same cell, and 3-D imaging make this ion microscopy modality a powerful tool in applied medical research and biology. A feasibility study on the use of SIMS for microscopic imaging of small animal heart slices is ongoing: SIMS could represent an important new methodology for microscopic imaging of coronary microcirculation. Coronary microcirculatory vasoconstriction has been documented during ischemia in patients with severe coronary stenosis. SIMS could offer the possibility to reveal suitable isotopic tracers of coronary flow under different conditions of perfusion pressure, as well as appropriate markers of ischemia. The effective use of ion microscopy for biological research requires reliable sample preparations and evaluation of SIMS matrix effects on image quantification. 8µm- coronal sections of rat heart previously included in OCT and frozen in liquid nitrogen were mounted on indium foils (backed on an aluminium block of 2,5 cm diameter and 3mm thickness) The preparations were moved on dry ice to a vacuum freeze drier where the sample remained for 2-5 hours. After the freeze drying phase, the samples gently reached room temperature. Adjacent cryo-sections were separately analysed with Scanning Electron Microscopy (SEM) and SIMS to correlate the morphological information and the quantitative imaging of ions. Preliminary result on cryogenic sample preparation and SIMS analysis of rat heart are reported.

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P12

BBiioorreesseeaarrcchh && PPaarrttnneerrss

ELIMINATION OF HEAVY METALS INTOXICATION. SEQUENTIAL MICRONUTRITION M. Lugand Bourgeois, A. Cornely Bioresearch and Partners – Route de l’Ile au Bois – 1870 Monthey (CH), Swizerland; E-mail-. bioresearch@bioresearch.ch By restoring homeostasis through the use of a nutritionnal new way, Bioresearch & Partners is to make a major contribution for the management of metal-induced toxicities, should they be punctual or chronic. The studied population focused on patients suffering from heavy metal-induced pathology. At the beginning and during the nutrition intervention period, patients were monitored for several other patho-physiological and psychosomatic symptoms as well as for blood hypersensitivity to metals Method Study: 6-month open trial Inclusion: 26 patients (18 to 75) with significant metal-induced symptoms: MELISA positive (Hg, Al, Ni, Au, Cd, Pd, Sn) blood test measuring hypersensitivity to metals that works by placing a range of metals into contact with T lymphocytes and monitoring the reaction as well as for several patho-physiological and psychosomatic symptoms fatigue, pain and inflammatory symptoms. six capsules per day Monitoring 3-6 months: patient patho-physiological and psychosomatic symptoms, Melissa test Result: between 3 -6 months, patients recorded a significant improvement in their pathological symptoms At 6 months: Fatigue (75 % of the patients before treatment): 90% reported significant reduction in asthenia. Gastro-intestinal symptoms (50% of the patients before treatment) symptoms disappeared in 70% of the patient population. Rhumatoid pain (60 % of the patients before treatment) symptoms disappeared in 50%of the patient population

- Stimulation of T lymphocytes by Hg, Pd, Al, Sn, and Au, in the MELISA assay, returned to normal values in all patients after 6 months,( Ni and Cd: 12 months).

Conclusion: elimination of symptoms and normalisation of the melisa test (lymphocytes reaction) offer a satisfactory response to the required therapeutic aims in a population presenting intoxication to heavy metals that can be used in monotherapy or in combination with other metal elimination approaches. BIORESEARCH & PARTNERS, BioArk, Route de l’Ile-au-Bois, 1870 Monthey, Switzerland - http://www.bioresearch.ch

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P13 AN EFFICIENT METHOD FOR THE CHARACTERIZATION OF THE INTERACTION OF PESTICIDES AND DIFFERENT SOIL SAMPLES

Csaba Csutorás, Attila Kiss Eszterházy Károly College, Department of Chemistry, Leányka str. 4., H-3300, Eger, Hungary

In our research group the investigation of the photodegradation of pesticides of different type is a traditional research topic. The mechanism of their photodegradation was tried to reveal exactly by the characterization of the degradation products. A special, immersable UV-light source emitting light of 254 nm effected degradation. Identification of degradation species was implemented by GC-MS technique.

Recently we started an extensive study for the investigation of the interaction of pesticides and their degradation products with different soil types. A suitable, simple model was created, similarly to column chromatography. The column was filled with the soil sample, and chloroform or ethyl acetate was used as eluent. Measuring the amount of eluent that is necessary to wash down the pesticide or degradation product from the column can deliver important data about the bond between the soil and pesticide. Hereby the environmental risks can be thoroughly revealed, not only for the pesticides but also for the photodegradation products. Atrazine, prometryne and terbutryne as representatives of s-triazine type pesticides, as well as diurone and carbendazime to represent different chemical structures were investigated. The polar or apolar character of the pesticide, or its degradation products seems to be the most important factor in the soil-pesticide interaction.

The research on revealing the exact reaction mechanisms of photolytic degradation of pesticides and the soil-pesticide interactions contribute to the proper understanding of behaviour of pesticides, and point out the possible environmental risk factors and ways of proper application.

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P14 DETERMINATION OF AL, Ca, Cu, Fe, Mg, Mn, P AND Zn IN POTATO AND GRAIN BY MEANS OF ETV-ICP-OES Albena Detcheva1, Juergen Hassler2 1Bulgarian Academy of Sciences, Institute of General and Inorganic Chemistry, Acad. G. Bontchev Str. Bl. 11, BG-1113 Sofia, Bulgaria 2 ESK Ceramics GmbH & Co. KG, P.O.Box 1526, D-87405 Kempten, Germany

The direct solid sampling spectroscopic methods based on electrothermal vaporization of powdered samples proved to be a good alternative to methods using wet digestion in sample preparation because of the simple sample pretreatment (if any), lower contamination risks, lower sample consumption and lower detection limits.

Electrothermal vaporization, combined with inductively coupled plasma optical emission spectrometry (ETV-ICP-OES) is a multielement technique, which is suitable for trace and minor element determination in different matrices. In the present work the ETV-ICP-OES was successfully applied for the determination of a number of elements in plant materials (potato and grain). The ETV-ICP-measurements were performed on a Thermo Jarrell Ash inductively coupled plasma spectrometer (model IRIS-AP) and an ETV-4000 combined with a 50-fold sampler (Spectral Systems).

Different calibration approaches were used for the determination of Al, Ca, Cu, Fe, Mg, Mn, P and Zn in plant tissues, using several types calibration standards - certified reference materials of biological origin, graphite-based home-made standards as well as dried aqueous standard solutions. The results were compared with those, obtained by an independent method – routine flame atomic absorption spectrometry after microwave acid digestion. There was a good agreement between the two data sets.

It was shown that the ETV-ICP-OES is a fast and reliable technique for trace and minor element determination in plant – materials like potato and grain. The metrological characteristics of the proposed method were discussed.

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P15 COMPOSITIONAL INVESTIGATION ON SETTLED AND AIRBORNE DUSTS WITH MINERALOGICAL METHODS Izabella Melinda Farkas1, Tamás G. Weiszburg2 1 Environmental Research Co-ordination Centre, Eötvös Loránd University, Pázmány Péter sétány 1/C, 1117 Budapest, Hungary; E-mail: farkasizabella@yahoo.com 2 Department of Mineralogy, Eötvös Loránd University, Pázmány Péter sétány 1/C, 1117 Budapest, Hungary; E-mail: weiszburg@ludens.elte.hu Solid contaminants get into the air as dusts, varying widely in both composition and size distribution. Dusts are classified into two groups upon grain size: airborne dust (< 10 µm) and settled dust (> 10 µm). Dust can be of natural or anthropogeneous origin. Natural dusts originate from soil erosion, forest fires, biogenic materials, sea salts above the sea surface, and volcanic eruptions. Anthropogeneous dust arises mainly from industrial activity and transportation. Phase (mineral) composition and size distribution of dusts establish the effect that dusts have on the environment and health. In Romania, county-level environment agencies collect settled dust samples on a monthly basis. The collected samples are weighted. In this work we studied settled dust samples that were collected from 18 localities in the period March–July 2003 by the Environment Protection Agency of Cluj County, Romania. Our main aim was to give the phase (mineral) composition of these dusts. The collected samples varied widely in particle diameter, so it was essential to separate them upon grain size. The following 7 grain size fractions have been set up: >400 µm, 400–250 µm, 250–125 µm, 125–63 µm, 63–32 µm, 32–1 µm and <1 µm (the first six fractions were obtained by sieving, while the last one by settling). Grain size distributions have been considered, too. Fractions above 63 µm have been studied under the stereomicroscope. Routine powder X-ray measurements have been carried out on the samples, while the most typical grains have been characterised by scanning electron microscopy (SEM equipped with EDX), too. Our results suggest that the minerals of the settled dust originate predominantly from soil erosion, accompanied by industrial contaminants. There is a strong correlation between grain size and composition: the proportion of mineral constituents increases with decreasing grain size. Among the identified constituents, the following have natural origin: quartz, CaCO3, feldspar, dolomite, mica, chlorite, anhydrite (?), goethite, lepidocrocite, kandite-group minerals, amphibole group, rutile, kianite, serpentine asbestos, ilmenite, halite, amphiboles and the biogenic materials like insects and plant fragments. Of anthropogenic origin are the silicium-carbide, corundum, cristobalite, tridymite, spinel-structure compounds and plastic materials. On the X-ray powder diffraction patterns micas, kandite-group minerals and halite occur mainly in the lower grain size fractions (63–1 µm, 63–32 µm, 32–1 µm). From the SiO2 polymorphs cristobalite is present in large amounts in some of the samples. This material may eventually get enriched in the respirable fraction, too, carrying increased health risk. Tridymite is subordinate relative to cristobalite, and occurs only in samples that contain silicium-carbide as well. In the case of settled dust special caution should be taken when drawing consequences on the geological background. In the case of a sample locality yielding salt-rich samples, the salt may well originate from a surface-exposed salt dome, however, contaminations from the winter salt treatments of the road cannot be excluded either. Another example is the sampling locality Turda, where the samples contained large amounts of calcite. Calcite may again come from the local outcropping limestones, however, the nearby cement factory, using calcite as base material for cement production, should be considered, too.

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P16 MANY-SIDED STRATEGY FOR SCREENING AND IDENTIFICATION OF UNKNOWN CONTAMINANTS IN WATER FOR HUMAN CONSUMPTION E. Ferretti, L. Lucentini, E. Veschetti, L. Bonadonna, A. Stammati, L. Turco, M. Ottaviani Istituto Superiore Sanità, Dipartimento Ambiente e connessa Prevenzione Primaria. Rome (Italy) In the European Union drinking water quality is currently assessed according to common standards described in the Directive 98/83/EC. Limit values for a definite number of parameters has been set up in order to establish the minimum requirements that water must meet when intended for human consumption. Unfortunately, routine methods for the determination of chemicals and microbiological agents listed in the Directive may fail to detect a lot of potential threatening agents that could intentionally be introduced in water supply systems within terrorism attacks. Moreover, the time necessary to carry out a complete survey does not fit quite well with this kind of emergencies. In this paper, we propose a new strategy for the screening and the identification of target and unknown contaminants in potentially contaminated drinking water samples. A wide-range of biological (viability and macromolecular synthesis of cultured cells), microbiological (e.g, early warning systems, presence/absence, etc.) and physico-chemical techniques (LC-UV/Vis, LC-IC, ICP-OES, GC-MS, LC-MS, etc.) are taken into account for the detection of different classes of contaminants (e.g, inorganic and organic compounds, metals, algal toxins, bacterial indicators of faecal contamination, enteric pathogens, staphylococci, actinomycetes, etc.). and of their effects. The proposed many-sided approach has been successfully applied in the research of mandatory parameters and unknown chemicals and microbiological agents in two potentially contaminated samples of drinking cooler waters.

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P17 ICP-OES DETERMINATION OF VANADIUM IN MICE TISSUES AFTER ORAL EXPOSURE E.Veschetti, B.Cittadini, M.Ottaviani, P.Leopardi, R.Crebelli Istituto Superiore Sanità, Dipartimento Ambiente e Connessa Prevenzione Primaria. Rome (Italy) Significant levels of vanadium have recently been detected in ground waters of active volcanic areas. Most of springs, drainage galleries and wells present on their surface are currently a valuable drinking water resource for a number of little and medium urban centres. Therefore, a growing interest about toxicological effects of vanadium exposure on consumers’ health has been shown in recent times. Toxicological studies currently available in literature proved that vanadium is responsible for a variety of biological effects as a result of the interaction with living cells. In-vitro experiments showed that low doses of vanadate ion induce DNA damage and chromosome malsegregation. On the other hand, insufficient data are available on in-vivo effects so that the characterization of the genotoxic hazard related to the oral intake has so far been uncertain. In order to provide a contribution in filling the knowledge gap, the authors are investigating micronuclei and DNA lesions in mice treated for 5 weeks with growing concentration (0.75 – 1,500 mg/) of vanadyl and vanadate. The results are also being related to vanadium concentration in mice tissues at the end of the treatment. For this end, a simple pre-treatment procedure for the digestion of small samples (0.07 – 0.70 g) of kidney, liver, spleen, bone and testis removed from sacrificed mice was developed. The procedure is based on acid (HNO3) digestion of each sample in 2-mL screw-capped plastic tubes heated from 50 to 65°C with 15-min steps and vented to room pressure after every step. In this article, a description of the applied procedure together with its performance compared to microwave acid digestion in Teflon vessels is provided. The implications of the experimental findings obtained for the assessment of the genotoxic risk of vanadium in drinking water are also discussed.

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P18 EXPERIMENTAL MICROCOSM MODEL FOR EVALUATION OF N2O AND CO2 PRODUCTION OF CULTIVATED SOIL Györgyi Kampfl, Krisztina Kristóf, György Heltai Department of Chemistry and Biochemistry, Szent István University, H-2103 Gödöllő For environmental protection reasons it is important to study the greenhouse gas emission from cultivated soil. Relatively few data can be found in the international literature and even less in Hungary concerning the emission of CO2 and NOx from agricultural soils. Therefore the effect of the factors (as water supply, the way of N-supply, the role of soil organisms and heavy metal contamination) controlling the accumulation of N2O and CO2 in soil and release to the atmosphere was studied in microcosm laboratory model experiments. Three experiments were conducted using closed glass vessels, brown forest soil, KNO3 fertiliser, maize residue (as organic additive) and water. The experiments lasted for several weeks in 2002 and 2003. During these periods the vessels were stored in a thermostat at 15 °C. The N2O and CO2 content of the head space gas samples were analysed by a gas chromatograph in each experiment. (The N2 and O2 content was investigated only in the first measurement.) Concerning the results in the first experiment after a one week preconditioning period changes were detected in both the N2O and CO2 productions. The effects of different treatments caused similar tendencies regarding the changes of gas concentrations in time. Unfortunately definite tendencies in the changes of N2 and O2 concentration could not reliably be detected by the applied GC method. In the second experiment the amount of the applied N-fertiliser and water was increased to make the concentration changes of the released gases more significant. The N-fertiliser was applied in dissolved form also to make the N-uptake of soil micro-organisms easier. In agreement with the preliminary expectations the N2O production increased in a well detectable way due to the different treatments. However the CO2 concentration decreased slightly. To confirm and explain the experienced phenomena in the third experiment the size of the used vessels and the amount of the N-fertiliser were increased keeping the other parameters constant. Similarly to the results of the second measurement series an increased N2O release was detected to the effect of nitrogen fertilisation and added organic matter. The changes of the gas concentrations in time showed similar tendencies compared with the results of the first experiment. The presence of the organic matter buffered the effect of the cadmium contamination i.e. the heavy metal additive did not restrict the microbial activity so much in the presence of maize residue than it did without organic matter. Although this experience also corresponded to the results published in literature, the overall rate of the N2O-N loss differed significantly from the published nitrogen losses of similar experiments. To explain and to confirm the results further studies were needed.

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P19 INVESTIGATION OF THE MECHANISM OF PHOTODEGRADATION OF DIFFERENT PESTICIDES Attila Kiss, Csaba Csutorás Eszterházy Károly College, Department of Biochemistry, Leányka str. 6., H-3300, Eger, Hungary

Major goal of our study was to investigate photolytic degradation of distinctive types of pesticides being frequently applied in practice. Relevance of the research is supported by the fact that few information is available on behaviour of pesticides used in agriculture, as well as reaction mechanism of photodegradation is not revealed yet.

A special, immersable UV-light source emitting light of 254 nm effected degradation. Subsequent to the irradiation of the sample the isolation of the degradation products was carried out by column-, and gas-chromatography. Identification of degradation species was implemented by mass spectrometry.

Each of the studied 7 distinctive pesticides (metribusine, simazine, acetochlor, carbendasime, diurone, atrazine, chlorpyriphos) of definitely different chemical structure exhibited definite liability to undergo photolytic decomposition. The degradation mechanisms of the mentioned pesticedes were entirely revealed, and in each case 5 or 6 major degradation species have been detected. Loss of thiomethyl, alkyloxy, alkyl and chloro groups might be regarded as typical ways of transformation.

We observed significant differences in the kinetic behaviour of the investigated pesticides by GC-monitoring. The kinetic curves of photolytic decomposition made it possible to compare photosensibility and reaction mechanisms of the studied pesticides.

The research on revealing the exact reaction mechanisms of photolytic degradation of pesticides contributes to the proper understanding of behaviour of pesticides, and points out the possible environmental risk factors and ways of proper application.

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P20 INTEGRATED APPROACH FOR ASSESSING ECOSYSTEM HEALTH Laura Mancini1, Paolo Formichetti1, Elio Pierdominici1, Anna Maria D’Angelo1, Marcello Iaconelli1, Stefano Fabiani1, Maria Elena Beltrami1, Stefania Marcheggiani1, Stefano Larsen1, Giorgio Pace 1, Lorenzo Tancioni2, Paola Bottoni1 1National Institute of Health, V.le Regina Elena 299, 00161 Roma, ITALY 2Università di Tor Vergata, Roma, ITALY As requested by the Water Framework Directive (WFD) different tools were used to investigate the ecological status of the lower course of Tiber river (central Italy). In five sites we applied the FFI (Fluvial Functional Index) for the general ecosystemic evaluation; Macroinvertebrate communities were studied through the Italian IBE index; usual physical-chemical and microbiological analysis were performed; toxicological analysis of sediments and the study of diatoms communities were also performed. Possible use of fish communities as bio-indicators is discussed. This study has allowed us to experiment new indexes and biological indicators and, at the same time evaluate the ecological status of the urban course of the Tiber river which is highly compromised due to excessive run-off of urban-polluted waters.

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P21 HEAVY METAL MONITORING IN PHASEOLUS VULGARIS IN TRANSYLVANIA C. Majdik1, Á. Pénzes2, I. Bartók1,M. Tosa1, P. Moldovan1, F. D. Irimie1, F. Kilár2

1Babes-Bolyai University of Cluj Napoca, Faculty of Chemistry and Chemical Engineering 2University of Pécs, Faculty of Science, Analytical Chemistry Department

The European Union has really strict norms, which should be observed. That is why the monitoring of the heavy metal content of the vegetables is very important. Heavy metals can enter into the food chain through the contaminated vegetables. During the research, we determined the heavy metal (Cr, Zn, Cu, Cd, Pb) content of the Phaseolus vulgaris. The common bean is wide spreaded in Transylvania and people consume it all year long. If people consume the polluted vegetables in high quantities, heavy metals can accumulate in the human organism and can cause several diseases. In this study, we monitored the heavy metal content of the Phaseolus vulgaris in 8 counties and 16 habitations in Transylvania. The determination was made from dry weight with HNO3 shattering by ICP-AES (Spectroflame FMD 07, Germany). The highest amount of the five heavy metals was measured in Covasna/Kovászna county- Racosul De Sus/Felsőrákos and in Cluj/Kolozs county- Baciu/Bács. Further investigation was tend to follow the translocation of the heavy metals from the seeds into the tissues of the new plant. We observed the whole amount of the examined metals translocated into the new generation’s tissues. References Cary E. E., Allaway W. H., Olson O. E., 1977, Control of chromium concentration in food plants.1. Absorption and translocation of chromium by plants. J. Agr. Food Chem. 25:300-304. Vara Prasad M. N., De Oliveira Freitas H. M., 2003, Metal hyperaccumulation in plants-Biodiversity prospecting for phytoremediation technology Sarret G., Vangrosveld J., Manceau A., Musso M., D’Haen J., Menthonnex J. J., Hazemann JL., 2001, Accumulation forms of Zn and Pb in Phaseolus vulgaris in the presence and absence of EDTA, Environ. Sci. Technol. 35:2854-2859.

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P22 OVERPRESSURED LAYER CHROMATOGRAPHIC DETERMINATION OF AFLATOXIN B1, B2, G1 AND G2 IN RED PAPRIKA Ágnes M. Móricz1, Zsuzsa Fatér2, Klára H. Otta1, Ernő Tyihák3

1 Department of Chemical Technology and Environmental Chemistry, L. Eötvös University, P.O.B. 32, Budapest 112, H-1518 Hungary (moricz_am@yahoo.com) 2Budapest Institute of the National Public Health and Medical Officer Service, Budapest, Váci út 174. H-1138 Hungary 3 Plant Protection Institute, Hungarian Academy of Sciences, Budapest, P.O.B. 102, H-1525 Hungary Aflatoxins, secondary metabolites of some Aspergillus moulds, may be contamination in food and feed e.g. cereals, oilseeds, cafe and spices like red paprika. The toxic effect of aflatoxins on human health is known therefore, the European Commission set maximum levels for their presence in foodstuffs. In red paprika, intended for human consumption, the maximum level of aflatoxin B1 is 5 �g/kg and of total aflatoxin (aflatoxin B1, B2, G1 and G2) is 10 �g/kg. There is European standard (EN 14123:2003) for determination of aflatoxins in paprika (HPLC method with post column derivatization and immunoaffinity column clean-up), which is adopted to Hungarian standard (MSZ EN 14123). Method was developed to measure the aflatoxins in red paprika, which performs the requirements of methods of analytical investigations of mycotoxins (CR 13505:1999). Previously the paprika was defating by Soxhlet extraction, with what largely the dye substances leave too. After this, the sample preparation was followed according to AOAC description (Kamimura et al.). Liquid extraction was used, the extract was cleaned on column, and the eluate was evaporated. The dissolved residue was applied for Overpressure layer chromatography (OPLC). The quantitative determination of separated aflatoxins was performed by using densitometric evaluation. References G.A. Payne, J.L. Richard, Mycotoxins: Risks in Plant, Animal, and Human Systems Council for Agricultural Science and Technology, Ames, Iowa, USA, Printed in the United States of America 2003 Kamimura H., Nishijima M. et al., Simple, rapid cleanup method for analysis of aflatoxins and comparison with various methods (1985) J. AOAC 68 458-461.

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P23 REDOX REACTIONS GOING ON IN WINES Z. Murányi, Szabolcs B.Tóth Eszterházy Károly College, Department of Chemistry, Leányka str. 4., H-3300 Eger, Hungary The oxidation processes going on while producing and storing wines are a vital determinant in terms of the quality of wines. The conditions of the production (reductive, oxidative) significantly affect the character of wines, the oxidation processes taking place during the storing of wines contribute to the ageing of wines and the deterioration in quality. The goal of our analysis was the extraction of compounds whose amount is closely related to the amount of the oxygen absorbed by wine; this way, they can be employed effectively to observe the oxidation processes. Regarding these compounds, the conditions are the following: 1, Their formation, (existence) is clearly the consequence of the oxidation. 2, The process is to be slightly affected by substances used for the treatment of the wine (such as the amount of sulphur dioxide). 3, The method applied for their quantitative determination has to be simple, fast and inexpensive; as it is supposed to be a routine work. Considering the options of several methods and the types of compounds, we have come to a decision to choose an organic and an inorganic compounds: namely aldehydes and iron. Their amount is essential in terms of wine quality as well. The analysis of aldehydes has been performed by SPME-GC method, the sampling from wine samples has been done by head-space analysis. The determination of the proportion of FeII and FeIII has been performed by a direct photometric method developed by us. We have chosen various types of wine samples, which following the “airing” and standing at different length of time, were analysed with respect to the chosen compounds.

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P24 INDUCTIVELY COUPLED PLASMA-SECTOR FIELD MASS SPECTROMETRIC DETERMINATION OF TRACE ELEMENTS IN THE PM10 AIRBORNE DUST FRACTION Gábor Muránszky1, Mihály Óvári2, Gyula Záray1,3,4,5 1Eötvös University Faculty of Sciences, Cooperative Research Centre of Environmental Sciences, H-1518 Budapest, P.O.Box 32. 2Eötvös University, Institute of Chemistry, Dept. of Chemical Technology and Environmental Chemistry, H-1518 Budapest, P.O.Box 32. 3Eötvös University, Institute of Chemistry, Dept. of Inorganic and Analytical Chemistry, H-1518 Budapest, P.O.Box 32. 4Joint Research Group of Environmental Chemistry of the Hungarian Academy of Sciences & Eötvös University, H-1518 Budapest, P.O.Box 32. 5Hungarian Satellite Centre of Trace Elements Institute to UNESCO, H-1518 Budapest, P.O.Box 32.

The aim of this study is, to give a survey about the trace elements concentration and their changes with time in the PM10 fraction of the Budapestian airborne dust. Our goal was to determine the platinum group metals (PGMs) concentrations as well, since the growing number of cars fitted with catalytic converters emit increasing amounts of PGMs, and their effects on the human body aren’t clear yet.

Monthly two (one and four day duration) PM10 aerosol samples were collected at the Széna-tér, Budapest. Sampling started in September 2004 and it is made continuously. We performed the elemental analysis of the PM10 samples collected from September 2004 to February 2005. The analysis was performed by ICP-SFMS from the solutions which were obtained by aqua regia digestion and water extraction. The measured elements were: Ag As, Ba, Be, Bi, Cd, Co, Cr, Cu, Fe, Ga, Mn, Mo, Ni, Pb, Pd, Pt, Rb, Rh, Sb, Se, Sr, Te, Tl, U, V, Zn.

On the analytical data we performed chemometric analysis as well to acknowledge which pollutants derive from the same source.

From the data collected during this half a year we got to the conclusion that there is no signignificant tendency in the change of the concentration in monthly steps. In the future we like to compare the average concentrations according to seasons and yearly whether we can find some tendency.

We compared the average concentrations measured during this half a year with other data published in the literature from big cities such as Buenos Aires, Vienna, Madrid.

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P25

DETERMINATION OF PLATINUM, PALLADIUM AND RHODIUM IN HUMAN URINE BY ELECTROTHERMAL VAPORIZATION INDUCTIVELY COUPLED PLASMA SECTOR FIELD MASS SPECTROMETRY (ETV-ICP-SFMS) Mihály Óvári1, Michaela Zeiner2, Ilse Steffan2, Gyula Záray3,4,5

1Eötvös University, Institute of Chemistry, Dept. of Chemical Technology and Environmental Chemistry, H-1518 Budapest, P.O.Box 32. 2University of Vienna, Inst. of Analytical Chemistry and Food Chemistry, A-1030, Währinger Str. 38., Austria 3Eötvös University, Institute of Chemistry, Dept. of Inorganic and Analytical Chemistry, H-1518 Budapest, P.O.Box 32. 4Joint Research Group of Environmental Chemistry of the Hungarian Academy of Sciences & Eötvös University, H-1518 Budapest, P.O.Box 32. 5Hungarian Satellite Centre of Trace Elements Institute to UNESCO, H-1518 Budapest, P.O.Box 32.

Platinum group metals are widely used in the catalytic converters of cars in order to decrease the CO, NOx, and hidrocarbon emission. Aim of this study was to investigate the uptake of the platinum group metals (PGM) by employees working on streets with very high traffic.

Urine samples taken from tram drivers working in the city centres of the two capitals (Vienna, Austria and Budapest, Hungary) were analysed. The tram drivers work on lines with very high traffic.

For the determination of palladium and rhodium, electrothermal vaporization inductively coupled plasma sector field mass spectrometry was used. Platium was determined by direct introduction into the ICP-MS of the 10-fold diluted urine samples. For the evaluation, the mean concentration values of platinum, palladium and rhodium related to creatinine were used.

In this work, the analytical results and the comparison between the two cities will be described and discussed.

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P26 SPECTROSCOPIC CONFIRMATION OF ELECTROCATALYTIC BEHAVIOR OF AMPEROMETRIC CARBOHYDRATE DETECTION ON COPPER ELECTRODE Lívia Nagy1, Géza Nagy2 1 Hungarian Academy of Sciences, Research Group for Chemical Sensors 2 University of Pécs, Faculty of Sciences Department of General and Physical Chemistry 7624 Pécs Ifjúság u.6.; E-mail: lnagy@ttk.pte.hu

Copper electrode based detector cell proved advantageous in solving different analytical tasks. They gained application in HPLC analysis of sugars, organic acids, nucleotides, amino acids and other analyte content of sample solution of different origin.

Interestingly copper working electrodes can be used in amperometric mode in cases, where noble metal and carbon electrodes fail to work properly.

As it is well known, the product of electrochemical oxidation of organic molecules such as mono- and disaccharides form an insulating layer passivating the electrode. In basic solution this electrode fouling effect does not show up when copper working electrode is employed in amperometric mode.

This validates that there are substantial differences in mechanism of electrochemical oxidation of these analytes proceeding at the different electrode surfaces.

While the electrode passivation can be the result of a direct electrode exchange between the metal surface and the adsorbed analyte, two different working principle can provide electrode passivation free detection:

- The amperometric signal can be the result of an oxidation involving the electrochemical dissolution of the electrode metal. - Electrocatalytic action of the oxide film coating the electrode surface also can avoid electrode fouling.

In our work the applicability of copper electrode based detector cell was investigated in details for the analysis of different saccharides and organic acids.

In the experiments to be discussed here controlled potential macroelectrolysis was performed in basic solution with measuring the electric charge passing through copper working electrode containing cell. Atomic absorption analysis and ion chromatographic measurements of the mediawere carried out after electrolysis. In agreement with the electric charge, formate was detected as major product of the electrolysis, and only traces of copper ions could be detected.

The findings indicate that electrocatalytic oxidation of the analyte provides the amperometric signal for the detection of the species examined.

The authors highly appreciate the financial support from the Hungarian Scientific

Research Foundation (OTKA T 037403 and T-049502).

References: L. Nagy, G. Nagy P. Hajós, Sensors and Actuators B 76 ( 2001) 494-499 L. Nagy,

R. Bátai, K. Horváth, P. Hajós , G. Nagy, J. Chromatogr. A (in press)

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P27 CHANGES OF BRAIN WATER AND ION CONCENTRATION IN EXPERIMENTAL HYPOXIC BRAIN OEDEMA IN RAT Monika Pásztor T1., Gábor Blazsó2

1Csongrád County Institute of State Public Health and Medical Officer Service, Hungary 2Department of Pharmacodynamics and Biopharmacy, University of Szeged, Hungary

Simultaneous occlusion of both common carotid arteries in rats produced cerebral oedema. The hypoxic brain oedema is associated with the damages of brain capillary system.

A close correlation is existed among the appearance of symptoms and the elevation of brain water and Na+ content and diminution of K+ content in the total brain tissue in time, indicating the development of cerebral edema due to ischemia.

The disturbance of the brain water electrolyte balance indicates the damages of permeability of the brain capillary system.

The Blood-brain barrier (BBB) is an endothelial permeability barrier present in capillaries and selectively limits the influx and efflux of a wide variety of solutes and substances between blood and brain. The BBB is a dynamic and regulated structure but its barrier function can be disrupted physiologically by means of absorbed of heavy metals.

For study of the pathogenesis of hypoxic brain oedema generated by bilateral carotid ligation. The changing of the brain water and electrolyte contents were examined in different parts of the brain.

The element concentrations were determined with a Perkin-Elmer 4100 (Bodenseewerk Perkin Elmer GmbH, Überlingen, Germany) atomic absorption spectrophotometer in air-acetylene flame or ETA.

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P28 CADMIUM AND ZINC IN HUMAN BRAIN SAMPLES Ivett Peltz-Császma1, Erzsébet Andrási1, Alexandra Lásztity1, Peter Brätter2 1 Department of Inorganic and Analytical Chemistry, Institute of Chemistry, Eötvös University, Budapest, Hungary 2 Department of Trace Elements in Health and Nutrition, Hahn-Meitner Institut für Kernforschung, Berlin, Germany

Cadmium is known to be a toxic element, while zinc, which is a highly similar chemical entity, is essential for life (activator/metallic cofactor of numerous enzymes: alcalic phosphatase, dehydrogenase, carboxy-peptidase, superoxide-dismutase; syntheses of proteins and nucleic acids; development of immune response). We determined the concentrations of these selected elements in human brain samples, to study their behavior in an organ exposed less to environmental effects. The sample collection consisted of 5 brain regions of 5 German control and 5 Alzheimer’s diseased patients. We applied microwave assisted and - for the enhancement of the accuracy of the results – also Parr-bomb digestions for sample dissolution. The cadmium content of the digested samples was determined by ETAAS (DL.: 30 ng/g, calculated for the original brain samples) and ICP-MS (DL.: 5 ng/g /Cd-111/, 12 ng/g /Cd-114/, calculated for the original brain samples). The zinc concentrations were measured by ICP-MS and NAA (solid sample analysis). The accuracy of the data was certified by CRMs (NBS SRM1577 Bovine liver, IAEA-359 Cabbage) digested with the samples. The results of the different techniques were in good agreement. The presence of cadmium in the brain samples is unambiguous. There was no matrix disturbance observed, blank measurements were appropriate. The cadmium level of the studied brain samples varied from the detection limit to 150 ng/g (dry weight), in some cases about 500 ng/g, with great diversity. While the concentrations of the essential zinc were in the range of 20 and 70 µg/g (dry weight).

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P29 DETERMINATION OF POTASSIUM ISOTOPE RATIOS BY TIMS IN BIOLOGICAL FLUIDS Enikő Tatár1,2, Klaus Mayer3, Tamás L. Török4, István Virág1, Victor G. Mihucz5, Gyula Záray1,2,5 1Department of Inorganic and Analytical Chemistry, L. Eötvös University, P.O. Box 32, H-1518 Budapest, Hungary 2Hungarian Satellite Centre of Trace Elements Institute to UNESCO, P. O. Box 32, H-1518 Budapest, Hungary 3European Commission- JRC, Institute for Transuranium Elements, P. O. Box 2340, 76125 Karlsruhe, Germany 4Dept. of Pharmacodynamics, Semmelweis University, H-1445 Budapest, Nagyvárad-tér 4, P.O.B.: 370, Hungary 5Joint Research Group of Environmental Chemistry of Hungarian Academy of Sciences & L. Eötvös University, P. O. Box 32, H-1518 Budapest, Hungary

Potassium is an important element in various physiological processes. The use of inductively coupled plasma mass spectrometry for the potassium isotope ratio determination encounters some difficulties because the potassium isotopes (m/z 39, 40 and 41) are obscured by the intense background peaks from Ar+ and ArH+. A solution for the determination of 40K isotope would be the use of He plasma, but in this case, special ICP MS instruments are required or the use of the TIMS technique is needed.

The objective of this study was the determination of the potassium isotope ratios in smooth-muscle cells of Guinea-pig (Taenia caeci). A “so-called” diluting fluid containing 0.018 mol/dm3 lanthanum and 0.01 mol/dm3 lithium in a 0.89 mol/dm3 nitric acid solution was used for the removal of potassium ions from the tissues. Four experimental sets were carried out. The muscle samples were equilibrated in this diluting fluid for 24 hours, after a 60 minute-long incubation in (modified) Krebs solutions followed by a 10 minute-long washing step in a lanthane containing sucrose solution in order to remove the intracellular ions before determinations. The potassium concentrations of the samples, determined by double focusing inductively coupled plasma mass spectrometry, were between 15 and 50 µg/cm3 and the calcium concentration was in the range of 0.04 – 3. 6 µg/cm3 depending on the type of the solutions used for the incubation of the muscle samples.

As the potassium isotope ratio determination by mass spectrometer in presence of calcium is not possible because of the isobaric interference, the calcium removal from the sample before the analysis was mandatory. For this purpose, an Amberlite IRC 748 chelating resin was used. For the method development, pooled samples were prepared by taking aliquots from the samples prepared with the same method. Two cm3 pool sample, neutralised with 25 % ammonia solution, was mixed with 1 gram resin. The resin was flushed with water and the final volume of “calcium-free” solution was 10 cm3.

The 41K/39K and 39K/40K isotope ratio analysis of the calcium free solutions was performed by thermal ionisation mass spectrometry using baking Ta single filament procedure.

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P30 TRACE ELEMENTAL CHARACTERIZATION OF EDIBLE OILS BY ICP-AES AND GFAAS Michaela Zeiner1, Iva Juranovic Cindric2, Ilse Steffan1

1Institute of Analytical Chemistry and Food Chemistry, University of Vienna, Währingerstraße 38, A-1090 Vienna, Austria 2Department of Analytical Chemistry, Faculty of Science, University of Zagreb, Strossmayerov trg 14, HR-10000 Zagreb, Croatia The determination of trace elements in edible oils is interesting for nutrition because of the metabolic role of metals in organisms, for adulteration detection and oil characterization. The quantification of the trace element content of oils was carried out using microwave digestion procedures followed by ICP-AES and GFAAS. The sensitivity of the ICP-AES technique was sufficient for the determination of Ca, Fe, Mg, Na, and Zn. Since the samples contained very low amounts of Al, Cu, Co, Cr, K, Ni, Mn, and Pb, these elements were measured by GFAAS. Differences of metal concentrations for edible oils can be used as an additional possibility for oil characterization, besides the analyses of the organic components.

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P31 PRECONCENTRATION AND GFAAS DETERMINATION OF VANADIUM IN MINERAL WATERS AND PHARMACEUTICALS Katalin Zih-Perényi, Alexandra Lásztity L. Eötvös University, Department of Inorganic and Analytical Chemistry, H-1518 Budapest 112, P.O. Box 32.; E-mail: perkat@freestart.hu

The determination of vanadium is difficult by the highly sensitive technique, ICP-MS, due to the interferences originating from spectral overlapping of polyatomic ions. Preconcentration of vanadium was investigated on a chelating functionalized cellulose, imino-diacetic acid ethyl cellulose (IDAEC). The sample was pressed through a minicolumn filled with 40 mg IDAEC, washed to separate from matrix components and finally eluted by HNO3 in a flow injection manner. The eluate was analyzed by graphite furnace atomic absorption spectrometry (GFAAS). About 20 fold preconcentration was achieved.The detection limit in solution is 0.081 µgL-1. In case of elements capable for changing valence it should be proved that there is not difference in sorption of the different oxidation states. By this method quantitative sorption can be obtained for both vanadium species commonly present in aerated conditions, namely V(IV) and V(V). In spite of that fact, while at 2 µgL-1 concentration the sorption was almost quantitative, at relatively high concentration (0,002 M) the V(V) ions could not be enriched on the IDAEC in any pH-es. This fact can be explained by the formation of inert izopolyacid or vanadate, which decomposes at higher temperature or longer reaction time. At low concentration V(V) could be reduced to V(IV) and so can be sorbed.

The optimum pH range is wide, between pH 3 and 7 the sorption is similar. In more basic conditions vanadium is not reactiv due to hypovanadate formation, V4O9

2-. This species was identified by potentiometric titration in 0,002 M solution. The stability constants of the V-IDAEC forms, which are characteristic for the strength of sorption, are evaluated also and estimated to be coherent with the considerable sorption at the lower (pH 3-5) pH-es.

The method was verified by analysis of a certified reference material with good results. The mineral waters analyzed were medicinal and commercial ones, the drugs were water-soluble materials. To prove the reliability of the method for mineral waters and pharmaceuticals, the recovery of spike additions was studied from that matrises. It was justified that the high TDS (total dissolved solids) generally do not influences the retention. To model the effect of humic and fulvic acids, citrate was spiked to the samples in different concentration. It was concluded that the enrichment is sensible for the presence of complexing agents even in 0,002 M. The sorption is depend on the concentration and kind of drug matrix. Working with real samples the influenced ones could be selected upon the basis of the recovery of a spike addition and should be diluted or analyzed by standard addition method.

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P32 INTERACTION BETWEEN ECOSYSTEMS AND HUMAN HEALTH. Giuliano Cecchi, Laura Mancini National Institute of Health, V.le Regina Elena 299, 00161, Roma, ITALY; E-mail: giulianocecchi@libero.it and lmancini@iss.it The study of Ecosystem Health is a relatively recent discipline that has already provided new insights into numerous aspects of environmental management. One of the most interesting fields of study is the one investigating the relationships between ecosystem and human heath. The ecosystem approach to human health stresses the importance of cultural and social values in shaping the concept of health, both at human and at ecosystem level. The negative effects of impoverished ecosystems on human beings has been reported on many occasions. Despite this, the correlation between impacted ecosystems and poor human health is somewhat troublesome to demonstrate and more research is needed to assess the import of ecosystem services from less developed to more developed countries.

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P33 ALKYLPHENOLS AND ALKYLPHENOL ETHOXYLATES IN WILD FISHES FROM THE RIVER TEVERE (CENTRAL ITALY): A PILOT STUDY Fulvio Ferrara1, Daniela Caronia1, Fabio Fabietti2, Nicoletta Ademollo2, Laura Mancini1, Enzo Funari1 1Istituto Superiore di Sanità, Dipartimento di Ambiente e Connessa Prevenzione Primaria 2Istituto Superiore di Sanità, Centro Nazionale per la Qualità degli Alimenti e dei Rischi Alimentari Keywords: alkylphenol, seafood, human health risk The research has focused the attention on the occurrence of alkylphenols (APs) in two fish species (Leuciscus cephalus and Angulla anguillae) form the Tiber River. This is the second largest river in Italy, it crosses industrial districts, agricultural and urbanised areas including the city of Rome, before flowing to the Tyrrhenian Sea. Three sampling sites were selected, in a non industrialized area approximately 100 km before the estuary, immediately before Rome and in the middle of the city. Preliminary results showed that both octyl (OPs) and nonylphenols (NPs), together with their mono and diethoxylate precursors, occurred in all the samples analysed. The highest values of APs were found in the samples from the middle of the city (OP, 1.4 ng/g; OP1E, 17 ng/g; OP2E, 1.4 ng/g; NP, 30 ng/g; NP1E, 0.5 ng/g; NP2E, 1.4 ng/g). These levels were in the same range reported in fish species from other European rivers. NP was always the AP detected at the highest level. Among OPs, octylphenol monoethoxylated was found at concentrations always higher than OP. The levels of APs found in the fish species of Tiber were sensibly lower than those in seafood from the Adriatic (Italian East Coast) and Tyrrhenian Sea (west coast).

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P34 FIRST DATA ON THE ECOLOGICAL CHARACTERIZATION OF THE RIVER ALBEGNA (TUSCANY, ITALY) Paolo Formichetti1, Paolo Andreani2, Tancioni Lorenzo3, Carlo Iacomini4, Laura Mancini1 1National Institute of Health, V.le Regina Elena 299, 00161 Roma, Italy; E-mail: mancini@iss.it 2Provincia di Viterbo, Viterbo, Italy 3Università di Tor Vergata, Roma, Italy 4APAT, Roma, Italy The River Albegna (Tuscany, central Italy) is one of the most interesting biotopes in Central Italy, with a high biodiversity in its upper course, a thermo-sulphurated inflow at the middle, and an estuary reaching many kilometres upstream. It flows in an area of considerable naturalistic interest which requires policy for conservation and, where necessary, of restoration. Data about a multi-parametric study on this river are reported. The investigation was based on macro-invertebrates communities, using IBE methodologies, and ichthyofauna; the FFI (Fluvial Functional Index) method was applied to evaluate the ecosystem quality. Additionally, physical-chemical and microbiological analyses (sulphite-reducing Clostridia) were performed in the water and sediments. The study of macroinvertebrate communities showed a degree of pollution increasing downstream and a certain anthropic stress in the final part of the river. The inflow of hot and salt rich water instead seem not influencing the survival of macroinvertebrates. Microbiological data showed a certain faecal contamination even if the auto-depurative capacities of the river seems not compromised. Chemical analyses confirmed the increasing downstream pollution as well as IFF. This study confirmed the naturalistic importance of the Albegna River, a natural ecosystem that will need only minor actions to protect and recover the more compromised parts of it.

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P35 MACROINVERTEBRATE AND AQUATIC MACROPYTHES AS INDICATORS OF FRESHWATER QUALITY Giorgio Pace, Paolo Formichetti, Elio Pierdominici, Laura Mancini National Institute of Health, V.le Regina Elena 299, 00161 Roma, Italy; E-mail: mancini@iss.it

The aim of this study is the analysis of the benthic macroinvertebrate and aquatic macrophytes communities as biological indicator of water quality, following the integrated approach framework as requested by the Water Frame Directive 2000/60/CE. Supporting these two analysis classical chemical analysis were performed. Benthic macroinvertebrates were studied following IBE index, an Italian standardized method for freshwater monitoring (D.Lgs. 152/99).

Considering that no standard methodology exist for the Macrophytes analysis in Italy, the French standard GIS index was performed, hence it is a first experimental approach. Two natural streams, Farfa and Treja (left and right tributary of Tiber river) and two artificial channels Pedicata and Ninfa were studied. These surface waters are in central Italy and are typical of the Mediterranean ecoregion. Farfa and Treja streams have a different macroinvertebrate community probably due to both different geological substrate and anthropic pressure. IBE values were higher in Farfa stream.

On the other hand, the analysis of aquatic macrophytes was limited by their scarce presence. In the two artificial channels benthic macroinvertebrates communities were unbalanced towards less sensible taxa, because of the typology of running waters, probably not adequate for sensitive taxa. In these channels, aquatic macrophytes were abundant giving low GIS scores, showing a condition of diffuse pollution due to high eutrophication. The use of bioindicators at different trophic levels give more detailed informations about the condition of aquatic environments highlighting the benefits of the integrated approach in a study of ecosystem health.

This study is a contribution to the application of WFD in Italy.

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P36 INTERACTION BETWEEN 2,2’-DISELENADIBENZOIC ACID AND SERUM ALBUMINS INVESTIGATED BY FLUORESCENCE SPECTROSCOPY

Liu Yi*, Yang Chang-Ying, Hou An-Xin, Qu Song-Sheng

Department of Chemistry, College of Chemistry and Molecular Sciences, Wuhan University, Wuhan 430072, People’s Republic of China

The binding of 2,2’-diselenadibenzoic acid to bovine serum albumin (BSA) and human serum albumin (HSA) was studied by using fluorescence spectroscopy. The measurement was performed in Tris-HCl buffer aqueous medium at pH=7.40. The quenching constant at 303 K was (3.277 ± 0.046) × 1013 L mol-1 s-1 for BSA, and (3.946 ± 0.002) × 1012 L mol-1 s-1 for HSA. Decreasing of quenching was observed in association with temperature increase. Our findings show that the observed binding constant is dependent on the ionic strength of the medium. It is said that the electrostatic interactions play a role in the binding of 2,2’-diselenadibenzoic acid to serum albumin, in addition to the hydrophobic association. The decrease of the linearity of S-V plot demonstrates reduced binding of ligand to the protein in the presence of anionic surfactants sodium dodecyl sulfate (SDS), which indicates that 2,2’-diselenadibenzoic acid most likely bind to the hydrophobic pockets within sub domain IIA of serum albumin, the same site for SDS. Keywords: 2,2’-diselenadibenzoic acid; BSA; HSA; binding; fluorescence; SDS *E-mail: prof.liuyi@263.net; Tel: +86 27 87218284; Fax: +86 27 68754067. Supported by Sino-Hungarian Scientific and Technological Cooperation Project and National Natural Science Foundation of China.

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LIST OF PARTICIPANTS 1. Abrankó László Corvinus University of Budapest Department of Applied Chemistry H-1118 Budapest, Villányi út 29-33 Tel.: +361 482 6363; fax.: +361 466 4272 E-mail: laszlo.abranko@uni-corvinus.hu 2. B. Tóth, Szabolcs Eszterházy Károly College Department of Chemistry, H-3300 Eger, Leányka út 4., Tel.: +36 36-520-471; fax: +36 36-520-471 E-mail: szabib@ektf.hu 3. Bálintné Kristóf Krisztina Szent István University Department of Chemistry and Biochemistry H-2103 Gödöllő, Páter K. u. 1 Tel.: +36 28-522-073; fax: +36 28-410-804 E-mail: - 4. Bandi Éva Pannonpower Holding Ltd. H-7632 Pécs, Edison u. 1. Tel.: +36 72-534-310; fax: +36 72-534-296 E-mail: kalorikus@pannonpower.hu 5. Bartók Imola Babeş-Bolyai University Department of Chemistry and Chemical Engineering RO-400028, Cluj Napoca Kolozsvár (Cluj – Napoca) Arany János u. 11., Tel.: +40 740 340916; fax: +40 26 4590818 E-mail: bartokimola@yahoo.com 6. Berlinger Balázs Fodor József National Center for Public Health Central Chemical Laboratory Department for Metal Analysis H-1450 Budapest, P.O. Box 22., Tel.: +36 1 476-1185; fax: +36 1 476-1374 E-mail: berlingerb@fjokk.hu 7. Bezur László University of Politechnics and Economical Sciences Department of General and Inorganic Chemistry H-1111 Budapest, Szt. Gellért tér 4. Tel.: +36 1 463-1178; fax: +36 1 463-3408

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E-mail: bezur@mail.bme.hu 8. Bottoni, Paola Istituto Superiore di Sanità Department of Environment and Health Prevention Unit of Aquatic Ecosystems and Bathing Water Quality Viale Regina Elena 299 00161 Rome Italy Tel.: +39 06 4990 2219; Fax: +39 06 4990 2999 E-mail: paola.bottoni@iss.it 9. Braun Mihály University of Debrecen Department of Inorganic and Analytical Chemistry H-4010 Debrecen, P. O. Box 21 Tel.: +36 52 512-900; ext.: 22304, Fax +36 52 489-667 E-mail: braun@tigris.unideb.hu 10. Campanella, Luigi Università degli Studi "La Sapienza" Piazzale Aldo Moro 5 00185 Roma Italy E-mail: luigi.campanella@uniroma1.it 11. Canepari, Silvia University of Rome "La Sapienza" Department of Chemistry Piazzale Aldo Moro 5 00185 Roma Italy E-mail: silvia.canepari@uniroma1.it 12. Capodaglio, Gabriele University of Venice Department of Environmental Science Dorsoduro 2137, 30123 Venice Italy E-mail: capoda@unive.it 13. Caroli, Sergio Istituto Superiore di Sanità Viale Regina Elena 299 00161 Rome, Italy Tel.: + 39 06 4990 2052; fax: + 39 06 4990 2366 E-mail: caroli@iss.it 14. Colombini, Maria Perla Dipartimento di Chimica e Chimica Industriale Università di Pisa

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via Risorgimento 35, 56126 Pisa, Italy tel.: +39 050 2219305 or +39 338 5944223; fax: +39 050 2219260 E-mail: perla@dcci.unipi.it 15. Csutorás Csaba Eszterházy Károly College Department of Chemistry, H-3300 Eger, Leányka út 4., Tel.: +36 36-520-471; fax: +36 36-520-471 E-mail: csuti@ektf.hu 16. de Boer, Jacob Netherlands Institute for Fisheries Research Animal Sciences Group Wageningen UR P.O. Box 68, 1970 IJmuiden The Netherlands E-mail: Jacob.deboer@wur.nl 17. Dobos Melinda Lithosphere Fluid Research Lab Department of Petrology and Geochemistry Eötvös University Budapest (ELTE) H-1117 Budapest, Pázmány Péter sétány 1/c Tel.: +(36) 1 209-0555 ext. 8340 E-mail: melinda.dobos@gmail.com 18. Ebdon, Les University of Luton Park Square Luton LU1 3JU Tel.: +44(0) 1582 489255; Fax: +44(0) 1582 489362 E-mail: Les.Ebdon@luton.ac.uk 19. Farkas Izabella Melinda Environmental Research Co-ordination Centre Eötvös Loránd University, H-1117 Budapest, Pázmány Péter sétány 1/C Tel.: +36-702114226 E-mail: farkasizabella@yahoo.com or fibella@freemail.hu 20. Ferretti, Emanuele Istituto Superiore di Sanità Department of Environment and Primary Prevention Viale Regina Elena, 299 00161 Rome – Italy Tel.: +39 06 49902553; Fax : +39 06 49902348 E-mail: repacqua@iss.it

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21. Focardi, Silvano University of Siena Department of Environmental Sciences "G. Sarfatti" Via Mattioli 4 53100 Siena Italy E-mail: focardi@unisi.it 22. Fodor Péter Corvinus University of Budapest Department of Applied Chemistry H-1118 Budapest, Villányi út 29-33 Tel.: +36 1 482-61-64; fax: +36 1 466-42-72 E-mail: fodor.peter@uni-corvinus.hu 23. Forte, Maurizio ARPA Lombardia Dipartimento di Milano - U.O. Agenti Fisici Via Juvara 22, 20129 Milano (Italy) Tel. +39 02-74872.264; fax: +39-02-7012.4857 E-mail: M.FORTE@arpalombardia.it 24. Fuoco, Roger University of Pisa Via Risorgimento, 35 56126 Pisa (Italy) Tel.:+39 0502219254; Fax +39 0502219260 E-mail: fuoco@dcci.unipi.it 25. Gergely Valéria Corvinus University of Budapest Department of Applied Chemistry H-1118 Budapest, Villányi út 29-33 Tel.: +36 1 482-61-64; fax: +36 1 466-42-72 E-mail: valeria.gergely@stud.uni-corvinus.hu 26. Halász Gábor Szent István University Department of Chemistry and Biochemistry H-2103 Gödöllő, Páter K. u. 1 Tel.: +36 28-522-073; fax: +36 28-410-804 E-mail: - 27. Hargitainé Tóth Ágnes College of Nyíregyháza Department of Chemistry H-4400 Nyíregyháza, Sóstói u. 31/b Tel.: +36 42 599 456; fax: - E-mail: tothag@zeus.nyf.hu

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28. Hassler, Jürgen ESK Ceramics GmbH & Co. KG Max-Schaidhauf-Strasse 25 D-87437 Kempten Tel.: +49 831-5618-144; fax: +49 831-5618-134 E-mail: juergen_hassler@t-online.de 29. Heltai György Szent István University Department of Chemistry and Biochemistry H-2103 Gödöllő, Páter K. u. 1 Tel.: +36 28-522-073; fax: +36 28-410-804 E-mail: heltai.gyorgy@mkk.szie.hu 30. Horváth Erzsébet University of Veszprém Department of Environmental Engineering and Chemical Technology H-8200 Veszprém, Egyetem utca 10. Tel.: +36 30-348-75-79, Fax: +36 88-62-4533 E-mail: elizabet@almos.vein.hu 31. Kápolna Emese Corvinus University of Budapest Department of Applied Chemistry H-1118 Budapest, Villányi út 29-33 Tel.: +36 1 482-61-64; fax: +36 1 466-42-72 E-mail: emese.kapolna@uni-corvinus.hu 32. Kilár Ferenc University of Pécs Institute of Chemistry Department of Analytical Chemsitry H-7624 Pécs, Ifjúság út 6. Tel.: +36 72 503-600; ext.: 4851, Fax: +36 72 501-518 E-mail: ferenc.kilar@ttk.pte.hu 33. Kiss Attila Eszterházy Károly College Department of Biochemistry and Molecular Biology H-Eger, 3300, Leányka u. 6. Tel.: 06-20-329-4478 E-mail: attkiss@ektf.hu 34. Kristóf János University of Veszprém Department of Analytical Chemistry H-8200 Veszprém, Egyetem utca 10, Tel.: +36 30-348-75-79; Fax: +36 88-62-4533 E-mail: kristof@almos.vein.hu

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35. Kvamme, Kristin Bergen University College Faculty of Engineering, P.O.Box 7030, N-5020 Bergen Tel.: +47 55 58 75 69; fax : +47 55 58 77 90 E-mail: Kristin.Kvamme@hib.no 36. Kvammen, Ove Bergen University College Faculty of Engineering P.O. Box 6030, N-5020 Bergen Tel.: +47 55587618 E-mail: Ove.Kvammen@hib.no 37. Lakatos János University of Miskolc Department of Chemistry, H-3515 Miskolc-Egyetemváros Tel.: +36 46-565111; ext. 13-73; fax: +36 46-565110 E-mail: mtasotak@gold.uni-miskolc.hu 38. Lengyel Attila University of Miskolc Cooperation Research Centre of Mechatronics and Material Science H-3515 Miskolc, Egyetemváros Tel.: 36 46 565 372; fax: 36 46 565 371 E-mail: meakkk.ig@uni-miskolc.hu 39. Lugand Bourgeois, Marie Bioresearch and Partners Route de l’Ile au Bois CH-1870 Monthey Swizerland Tel.: +41 (0) 244736250; fax: +41 (0) 244736259 E-mail: bioresearch@bioresearch.ch or mlugand@bioresearch.ch 40. Maloschik Erik Plant Protection Institute of Hungarian Academy of Sciences H-1525 Budapest PO Box 102 Tel.: +36 1 3918665; fax: +36 1 3918655 E-mail: emal@julia-nki.hu 41. Mancini, Laura Italian National Institute of Health Viale Regina Elena 299 00161 Rome, Italy Tel.: +39 0649902773; fax: +39 0649902861 E-mail: lmancini@iss.it

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42. Marcus, Peter Ben-Gurion University of the Negev Environmental Analytical Laboratory (EAL) Rotem Industrial Park, Mishor Yamin ISRAEL 86800 Tel.: +972 8-6571331; Fax +972 8-6571332 E-mail: eal@bgumail.bgu.ac.il 43. Margitai Zita University of Debrecen Department of Inorganic and Analytical Chemistry H-4010 Debrecen, P. O. Box 21 Tel.: +36 52 512-900; ext.: 22304, Fax +36 52 489-667 E-mail: margitaiz@freemail.hu 44. Mihucz Viktor Gábor Joint Research Group of Environmental Chemistry of the Hungarian Academy of Sciences & L. Eötvös University H-1518 Budapest, P.O. Box 32 Tel: +36 1 209055 ext. 1223; fax: +36 1 372 2608 E-mail: vgmorales77@yahoo.com 45. Móricz Ágnes L. Eötvös University Department of Chemical Technology H-1518 Budapest, P.O. Box 32 E-mail: moricz_am@yahoo.com 46. Nagy Géza University of Pécs Department of General and Physical Chemistry H-7624 Pécs, Ifjúság u.6. Tel./fax: +36 72- 503 600 E-mail: g-nagy@ttk.pte.hu 47. Nagy Lívia Research Group for Chemical Sensors of the Hungarian Academy of Sciences H-7624 Pécs, Ifjúság u.6 Phone/Fax: +36 72- 503 600 E-mail: lnagy@ttk.pte.hu 48. Ocskay Rita L. Eötvös University Department of Chemical Technology H-1518 Budapest, P.O.Box 32 E-mail: ocskayrita@freemail.hu 49. Osán János

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KFKI Atomic Energy Research Institute P.O. Box 49, H-1525 Budapest E-mail: osan@sunserv.kfki.hu 50. Óvári Mihály L. Eötvös University Department of Chemical Technology H-1518 Budapest, P.O. Box 32 Tel: +36 1 209 05 55, ext. 1114; fax: +36 1 372 26 08 E-mail: ovari@para.chem.elte.hu 51. Paksy László University of Miskolc Department of Inorganic Chemistry H-3515 Miskolc, Egyetemváros Tel.: 36 46 565 372; fax: 36 46 565 371 E-mail: akmpl@gold.uni-miskolc.hu 52. Papp István University of Debrecen Department of Mineralogy and Geology H-4032 Debrecen, Egyetem tér 1. Tel.: +36 52 512-900; ext. 22304; Fax: +36 52 489-667 E-mail: papis@delfin.unideb.hu 53. Pásztor Turák Monika Csongrád County Institute of State Public Health and Medical Officer Service H-6726 Szeged, Derkovits fasor 7-11. Tel: +36 62 592-557 E-mail: pastum@freemail.hu 54. Patriarca, Marina Istituto Superiore di Sanità Viale Regina Elena 299 00161 Rome Italy E-mail: m.patria@iss.it 55. Peltz – Császma Ivett Eötvös University, Department of Inorganic and Analytical Chemistry, H-1518 Budapest, P.O. Box 32. E-mail: csivett_peltz@yahoo.com 56. Pénzes Ágota University of Pécs Department of General and Physical Chemistry H-7624 Pécs, Ifjúság u.6. Tel./fax: +36 72- 503 600 E-mail: a_penzes@yahoo.com

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57. Pepponi, Giancarlo ITC-irst via Sommarie 18 38050 Povo, Trento, Italy E-mail: pepponi@itc.itű 58. Pietrodangelo, Adriana National Research Council Institute for Atmospheric Pollution Via Salaria Km. 29.300 CP 10 - 00016 Monterotondo Stazione - ROMA (Italy) Tel.: +39 6-90672263; fax: +39 6-90672660 E-mail: pietrodangelo@iia.cnr.it 59. Polyák Klára University of Veszprém Department of Earth and Environmental Sciences H-8200 Veszprém, Egyetem u. 10 Tel.: +36 88 624 832, fax: + 36 88 624 454 E-mail: polyak@almos.vein.hu 60. Posta József University of Debrecen Department of Inorganic and Analytical Chemistry H-4032, Debrecen, Egyetem tér 1. Tel.: +36 52 512-900; ext. 22427; fax: +36 52 489-667 E-mail: postaj@tigris.unideb.hu 61. Rezić, Iva University of Zagreb Faculty of Textile Technology Laboratory of Analytical Chemistry Tel.: +38 5981784504 E-mail: iva_rezic@net.hr 62. Righetti, Benito Ufficio Scientifico dell’Ambasciata d’Italia in Ungheria H-1143 Budapest, Stefánia út. 95. Tel./fax: + 36 1 3841760 E-mail: righetti@ambitalia.hu 63. Roda, Aldo University of Bologna Dept. of Pharmaceutical Sciences Via Belmeloro 6, 40126 Bologna, Italy Tel./fax: +39 051 343398 E-mail: aldo.roda@unibo.it

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64. Sabbioni, Enrico European Commission - Joint Research Center Institute for Health and Consumer Protection (IHCP) ECVAM Unit 21020 Ispra, Varese (Italy) Tel.: +39 0332 786342; fax: +39 0332 785994 65. Salma Imre Eötvös University Department of Chemical Technology and Environmental Chemistry, H–1518 Budapest, P.O. Box 32, Hungary E-mail: salma@para.chem.elte.hu 66. Sánta Imre University of Pécs Institute of Physics Department of General Physics and Laser Spectroscopy H- 7624 Pécs, Ifjúság u. 6. Tel: +36-72-501559; fax: +36-72-501571 E-mail: santa@fizika.ttk.pte.hu 67. Schäfer, Thomas University of Pisa Department of Chemsitry and Chemical Engineering Via Risorgimento, 35 56126 Pisa (Italy) Tel.: +39 0502219254; fax: +39 0502219260 E-mail. tom@ifc.cnr.it 68. Schäffer Richard Corvinus University of Budapest Faculty of Food Science Department of Applied Chemistry H-1118 Budapest, Villányi út 29-33 Tel.: +361 482 6253; fax.: +361 466 4272 E-mail: richard.schaffer@uni-corvinus.hu 69. Sörös Csilla Corvinus University of Budapest Faculty of Food Science Department of Applied Chemistry H-1118 Budapest, Villányi út 29-33 Tel.: +361 482 6253; fax.: +361 466 4272 E-mail: csilla.soros@uni-corvinus.hu 70. Stefánka Zsolt Hungarian Academy of Sciences Institute of Isotopes Department of Radiation Safety Konkoly-Thege Miklós út. 29-33.

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H-1121 Budapest Tel.: 0036-1-3923339; fax: 0036-1-3922533 71. Szlepák Emőke Szent István University Department of Chemistry and Biochemistry H-2103 Gödöllő, Páter K. u. 1 Tel.: +36 28-522-073; fax: +36 28-410-804 E-mail: szlepak.emoke@mkk.szie.hu 72. Tatár Enikő L. Eötvös University Department of Inorganic and Analytical Chemistry H-1518 Budapest, P.O. Box 32 Tel.: +36 1 209 0555, ext. 1209; fax: +36 1 372 2608 73. Üveges Márta Corvinus University of Budapest Faculty of Food Science Department of Applied Chemistry H-1118 Budapest, Villányi út 29-33. Tel.: +36 1 482 6253, Fax: +36 1 466 4272 E-mail: marta.uveges@uni-corvinus.hu 74. Varga Zsolt Hungarian Academy of Sciences Institute of Isotopes Department of Radiation Safety Konkoly-Thege Miklós út. 29-33. H-1121 Budapest Tel: +36-1-3923339; Fax: +36-1-3922533 E-mail: varga@iki.kfki.hu 75. Varga Imre Eötvös Loránd University Institute of Chemistry H-1518 Budapest P.O.Box 32, Hungary E-mail: vargaip@para.chem.elte.hu 76. Veschetti, Enrico Istituto Superiore di Sanità Department of Environment and Primary Prevention Viale Regina Elena, 299 – 00161 Rome – Italy Tel.: +39 06 49902553; Fax : +39 06 49902348 E-mail: repacqua@iss.it 77. Virág István L. Eötvös University Department of Inorganic and Analytical Chemistry H-1518 Budapest, P.O. Box 32

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Tel.: +36 1 209 0555, ext. 1315; fax: +36 1 372 2608 E-mail: agysejt@yahoo.com 78. Yao, Jun School of Environmental Studies China University of Geosciences, Wuhan, Hubei province, 430074, P. R. China Tel.:+86 27-67883036; fax:+86 27-67885032 E-mail: junyao@cug.edu.cn 79. Yi, Li Wuhan University College of Chemistry and Molecular Sciences Department of Chemistry, Wuhan 430072, People’s Republic of China 80. Záray Gyula L. Eötvös University Department of Inorganic and Analytical Chemistry H-1518 Budapest, P.O. Box 32 Tel.: +36 1 209 0555, ext. 1513; fax: +36 1 372 2608 E-mail: zaray@ludens.elte.hu 81. Zeiner, Michaela University of Vienna Faculty of Chemistry Institute of Analytical and Food Chemistry Waehringerstrasse 38, 1090 Vienna, Austria Tel: +43 1-4277-52366; fax: +43 1-4277-9523 82. Zihné Perényi Katalin L. Eötvös University Department of Inorganic and Analytical Chemistry H-1518 Budapest, P.O. Box 32 E-mail: perkat@freestart.hu